http://2012.igem.org/wiki/index.php?title=Special:Contributions&feed=atom&limit=20&target=Lucho.Moro&year=&month=2012.igem.org - User contributions [en]2024-03-29T00:03:13ZFrom 2012.igem.orgMediaWiki 1.16.0http://2012.igem.org/Team:Buenos_Aires/Results/DevicesTestingTeam:Buenos Aires/Results/DevicesTesting2012-10-27T04:05:13Z<p>Lucho.Moro: /* Results */</p>
<hr />
<div>{{:Team:Buenos_Aires/Templates/menu}}<br />
<br />
<br />
= Devices testing =<br />
<br />
== Experimental Setup ==<br />
<br />
=== Plasmids and BBs ===<br />
<br />
In order to construct the yeast expression plasmids we choosed 3 vectors, 2 with a tryptophan marker and 1 with an histidine marker:<br />
# '''pCM182/5''', which are centromeric plasmids with TRP1 marker, and with a doxycycline repressible promoter [Gari et al 1996]. <br />
# '''pEG202''', with a 2 ori, HIS3 marker and a constitutive promoter (PADH1). <br />
<br />
<br />
{| style="width:100%"<br />
|[[File:BsAs2012-plasmid-PEG202.jpg|400px]]<br />
|[[File:BsAs2012-plasmid-BPCM185.gif|340px]]<br />
|}<br />
<br />
The cloning we did was:<br />
<br />
{| class="wikitable"<br />
|<br />
! scope="row" style="background: #7ac5e8"|<partinfo>BBa_K792009</partinfo> (PoliHa)<br />
! scope="row" style="background: #7ac5e8"|<partinfo>BBa_K792010</partinfo> (TRPZipper2)<br />
! scope="row" style="background: #7ac5e8"|<partinfo>BBa_K792011</partinfo> (PoliHb)<br />
! scope="row" style="background: #7ac5e8"|<partinfo>BBa_K792012</partinfo> (PoliWb)<br />
|-<br />
! scope="row" style="background: #81BEF7"|pCM182 (TRPa)<br />
|<br />
! scope="row" style="background: #01DF01"|X<br />
|<br />
! scope="row" style="background: #01DF01"|X<br />
|-<br />
! scope="row" style="background: #81BEF7"|pCM185 (TRPb)<br />
|<br />
! scope="row" style="background: #01DF01"|X<br />
|<br />
! scope="row" style="background: #01DF01"|X<br />
|-<br />
! scope="row" style="background: #81BEF7"|pEG202 (HIS)<br />
! scope="row" style="background: #01DF01"|X<br />
|<br />
! scope="row" style="background: #01DF01"|X<br />
|<br />
|}<br />
<br />
<br />
==== Cloning protocol ====<br />
<br />
# Digestion of plasmids and TRP/HIS export device:<br />
##pCM182; pCM 185; BBa_K792010 y BBa_K792012 were digested with BamHI and Pst1 restriction enzymes.<br />
##pEG202; BBa_K792009 y BBa_K792011 were digested with BamHI and Not1.<br />
# Purification of digested vectors (pCM185; pCM182; pEG202) <br />
# Ligation of vectors and devices according the anterior table (T4 ligase protocol, overnight)<br />
# Transformation of E. Coli DH5a with the ligation products. Bacterias were plated on LB-agar with Ampicillin, and incubated over night at 37 °C.<br />
# Colonies were used for liquid cultures (LB + Ampicillin) and minipreps were made.<br />
# Constructions (vector + insert) were checked by digestion with restriction enzymes, and 1%-agarose gel (1kb and 100bp as markers).<br />
<br />
=== Yeast strains ===<br />
<br />
Once obtained the desired constructions, we transformed yeast strains:<br />
# TCY3081: W303, bar1-, ura3::PAct1-YFP<br />
# TCY3128: W303, bar1-, leu2:: Pprm1-CFP 405<br />
<br />
<br />
<br />
=== Final transformed strains ===<br />
<br />
We got the following '''transformed strains''':<br />
<br />
{|<br />
|<br />
{| style="width:50%"<br />
| rowspan="3" | [[File:BsAs2012-icono-Cepa1.jpg | 200px]]<br />
! scope="row" style="background: #7ac5e8"| Name<br />
| '''YFP_TRPa_TRPZipper2'''<br />
|-<br />
! scope="row" style="background: #CEE3F6"|Strain<br />
|TCY3081 (YFP)<br />
|-<br />
! scope="row" style="background: #CEE3F6"|Plasmid<br />
|pCM182 (TRPa) + <partinfo>BBa_K792010</partinfo> (TRPZipper2)<br />
|}<br />
|<br />
{| style="width:50%"<br />
| rowspan="3" | [[File:BsAs2012-icono-Cepa2.jpg | 200px]]<br />
! scope="row" style="background: #7ac5e8"| Name<br />
| '''YFP_TRPa_PoliWb'''<br />
|-<br />
! scope="row" style="background: #CEE3F6"|Strain<br />
|TCY3081 (YFP)<br />
|-<br />
! scope="row" style="background: #CEE3F6"|Plasmid<br />
|pCM182 (TRPa) + <partinfo>BBa_K792012</partinfo> (PoliWb)<br />
|}<br />
|}<br />
<br />
{|<br />
|<br />
{| style="width:50%"<br />
| rowspan="3" | [[File:BsAs2012-icono-Cepa3.jpg | 200px]]<br />
! scope="row" style="background: #7ac5e8"| Name<br />
| '''YFP_TRPb_TRPZipper2'''<br />
|-<br />
! scope="row" style="background: #CEE3F6"|Strain<br />
|TCY3081 (YFP)<br />
|-<br />
! scope="row" style="background: #CEE3F6"|Plasmid<br />
|pCM185 (TRPb) + <partinfo>BBa_K792010</partinfo> (TRPZipper2)<br />
|}<br />
|<br />
{| style="width:50%"<br />
| rowspan="3" | [[File:BsAs2012-icono-Cepa4.jpg | 200px]]<br />
! scope="row" style="background: #7ac5e8"| Name<br />
| '''YFP_TRPb_PoliWb'''<br />
|-<br />
! scope="row" style="background: #CEE3F6"|Strain<br />
|TCY3081 (YFP)<br />
|-<br />
! scope="row" style="background: #CEE3F6"|Plasmid<br />
|pCM185 (TRPb) + <partinfo>BBa_K792012</partinfo> (PoliWb)<br />
|}<br />
|}<br />
<br />
{|<br />
|<br />
{| style="width:50%"<br />
| rowspan="3" | [[File:BsAs2012-icono-Cepa6.jpg | 200px]]<br />
! scope="row" style="background: #7ac5e8"| Name<br />
| '''CFP_HIS_PoliHa'''<br />
|-<br />
! scope="row" style="background: #CEE3F6"|Strain<br />
|TCY3128 (CFP)<br />
|-<br />
! scope="row" style="background: #CEE3F6"|Plasmid<br />
|pEG202 (HIS) + <partinfo>BBa_K792009</partinfo> (PoliHa)<br />
|}<br />
|<br />
{| style="width:50%"<br />
| rowspan="3" | [[File:BsAs2012-icono-Cepa5.jpg | 200px]]<br />
! scope="row" style="background: #7ac5e8"| Name<br />
| '''CFP_HIS_PoliHb'''<br />
|-<br />
! scope="row" style="background: #CEE3F6"|Strain<br />
|TCY3128 (CFP)<br />
|-<br />
! scope="row" style="background: #CEE3F6"|Plasmid<br />
|pEG202 (HIS) + <partinfo>BBa_K792011</partinfo> (PoliHb)<br />
|}<br />
|}<br />
<br />
<br />
<br />
== Secretion Rate of Trp as a function of culture growth ==<br />
<br />
The first step was to actually check if the construct works: do the transformed yeast strains - with the tryptophan devices- actually secrete tryptophan into the medium?<br />
<br />
To test this we used the following strains:<br />
<br />
{|<br />
|<br />
{|<br />
|-<br />
|[[File:BsAs2012-icono-Cepa3.jpg|200px]]<br />
|[[File:BsAs2012-icono-Cepa4.jpg|200px]]<br />
|[[File:BsAs2012-icono-YFP-185.jpg|200px]]<br />
|- align="center"<br />
|YFP_TRPb_TRPZipper2<br />
|YFP_TRPb_PolyWb<br />
|YFP_TRPb<br />
|}<br />
| rowspan="2" |<br />
{|<br />
| [[File:BsAs2012-icono-YFP.jpg | 200px]]<br />
|- align="center"<br />
|YFP<br />
|}<br />
<br />
|-<br />
|<br />
{|<br />
|-<br />
|[[File:BsAs2012-icono-Cepa1.jpg|200px]]<br />
|[[File:BsAs2012-icono-Cepa2.jpg|200px]]<br />
|[[File:BsAs2012-icono-YFP-182.jpg | 200px]]<br />
|- align="center"<br />
|YFP_TRPa_TRPZipper2<br />
|YFP_TRPa_PolyWb<br />
|YFP_TRPa<br />
|}<br />
<br />
|}<br />
<br />
<br />
===== Protocol =====<br />
<br />
*We started 5ml cultures with 3 replica until they reached exponential phase, overnight, using a -T medium.<br />
*Starting OD for the assay 0.1 (exponential phase). <br />
*We measured OD every hour until they reached an OD: 0.8 (5 hs approximately). <br />
*We measured the Trp signal for each culture medium using the spectrofluorometer.<br />
<br />
NOTE: The fluorescence meausurements taken for the Tryptophan in the medium, take into account both the aminoacid secreted by the device and the one diffused. <br />
<br />
Therefore the secretion rates calculated will be higher than the actual ones. We used an empty plasmid as control to study tryptophan diffusion.<br />
<br />
We used a simple model to measure the secretation rate for all the strains. Since the cultures are in exponential phase, we take<br />
<br />
[[File:BsAs2012-eqTrp1.jpg | 225px]]<br />
<br />
After a few calculations, we find that<br />
<br />
[[File:BsAs2012-eqTrp2.jpg | 225px]]<br />
<br />
===== Results =====<br />
<br />
Next we show the average OD for each strains used, needed to calculate the export rates of Trytophan.<br />
<br />
{|<br />
[[File:BsAs2012Odvstimea4.jpg | 250px]]<br />
|<br />
[[File:BsAs2012Odvstimeab.jpg | 250px]]<br />
|}<br />
Figure1. Cell density vs time for different strains in media that lack tryptophan or in synthetic defined medium (SC).<br />
<br />
We can see the cell growth for strain YFP transformed with the tryptophan devices in media that lack tryptophan (-H), compared with the strain transformed with empty plasmid in rich Synthetic Define medium (SC). This last strain grew faster than the others, we again see that our strains needs a longer amount of time to grow. <br />
<br />
[[File:BsAs2012rate2.jpg| 450px]]<br />
<br />
Figure2. Tryptophan export rate for different tryptophan devices or with empty plasmids.<br />
<br />
We can see that YFP strain transformed with tryptophan export devices hace higher secretion rates, compared with the YFP yeast transformed with empty plasmid that should only diffuse tryptophan through the membrane. <br />
This negative control is about 10% of secretion rate found for the strains with devices and is the same order as the error associated.<br />
<br />
== Tryptophan secretion at increasing histidine concentrations ==<br />
<br />
We asked ourselves which was the dependance of tryptophan secretion on the amount of histidine in medium. We carried on this test in order to determine whether the secretion of tryptophan depends on the concentration of another aminoacid in the media, such as histidine and what would be the necessary amount of histidine in medium for the start of our system. <br />
<br />
In this experiment we used strains: <br />
<br />
<br />
{|<br />
|-<br />
|[[File:BsAs2012-icono-Cepa3.jpg|200px]]<br />
|[[File:BsAs2012-icono-Cepa4.jpg|200px]]<br />
|[[File:BsAs2012-icono-YFP-185.jpg|200px]]<br />
|- align="center"<br />
|YFP_TRPb_TRPZipper2<br />
|YFP_TRPb_PolyWb<br />
|YFP_TRPb (control)<br />
|}<br />
<br />
<br />
<br />
<br />
<br />
<br />
<br />
===== Protocol =====<br />
<br />
# Starters of each strain used were grown over night in -T media, at 30 °C in shaker.<br />
# After 12 hs, cells were pelleted and washed with -HT media.<br />
# Cultures with increasing concentrations of histidine (0X, 1X, 1/4X and 1/16X) were set at an initial OD: 0.1, for each strain with 2 replica.<br />
# We left the cultures in shaker at 30ºC for 5 hours. After that time, we measured the final OD reached by cultures with the use of a spectrophotometer and the amount of tryptophan present in medium with a spectrofluorometer.<br />
<br />
===== Results =====<br />
<br />
[[File: BsAs2012TrpvHis1.jpg|350px]]<br />
<br />
Figure 3. Final tryptophan concentration as a function of histidine concentration in the media.<br />
<br />
<br />
With this result we can infer that tryptophan export has no clear dependence on histidine concentration in the media. Therefore, supporting the assumption of the model.<br />
<br />
== Measurement of Trp in medium and Basal Production ==<br />
<br />
To check the efectiveness of our biobricks, we must first determine the ammount of tryptophan secreted by natural strains to the medium, so we can compare. With that end in mind, we designed a protocol for measurement of tryptophan in medium, based in its fluorescense at 350nm, when excited with 295nm light.<br />
As a previous step, we checked that none of the other aminoacids used in the medium interferes, by graphically comparing the spectres for uncomplemented medium and medium complemented with leucine, uracile and histidine, at an appropiate range.<br />
<br />
To determine Trp concentration, we must first have a way to transform our readings (intensity) to a more useful output, so we made a calibration curve, through serialized 1:2 dilutions of our medium, which Trp's concentration is 50mg/mL, until approximately constant intensity.<br />
<br />
The procedure to measure secretion rates will be growing the strain from a known OD in exponential growth phase in -T medium and plotting it's OD over time, spin-drying at time=t, retrieving the supernatant's Trp concentration and dividing it by the integral of OD vs. time between time=0 and time=t, so we get to a rate which will be proportional to the number of cells in the culture, which means we can actually compare between different strains. Since our medium is free from Trp, all of it should come from within the cells, and if the culture is growing at exponential rates, lysis should be negligible, so the only explanation would be cells exporting their own Trp.<br />
<br />
<br />
{| class="wikitable" border="1"<br />
|-<br />
|<!--column1-->[[File:Trp-bsas2012.png | 250px]]<br />
|-<br />
|Graph:Tryptophan calibration curve<br />
|}<br />
<br />
<br />
<br />
==== Results ====<br />
<br />
As can be seen from the graph the screening of the concentration of the Trp in medium describes an almost lineal function. Through this experiment we can be sure that we would be able to measure increase of Trp in medium as it is exported from the cells, within the biological range of export.<br />
The sensitivity of this method seems to be enough to detect concentrations as low as ~0.02mg/mL, and as high as 50mg/mL, maybe more. Since our medium is 50mg/mL, we assume that's the saturation point of the curve. If we get bigger intensities than the one corresponding to it, we will dilute the sample.<br />
<br />
Because of time constraints, we haven't been able to check the method with either our designed strains nor the non-exporting ones.</div>Lucho.Morohttp://2012.igem.org/Team:Buenos_Aires/Results/DevicesTestingTeam:Buenos Aires/Results/DevicesTesting2012-10-27T04:01:09Z<p>Lucho.Moro: /* Results */</p>
<hr />
<div>{{:Team:Buenos_Aires/Templates/menu}}<br />
<br />
<br />
= Devices testing =<br />
<br />
== Experimental Setup ==<br />
<br />
=== Plasmids and BBs ===<br />
<br />
In order to construct the yeast expression plasmids we choosed 3 vectors, 2 with a tryptophan marker and 1 with an histidine marker:<br />
# '''pCM182/5''', which are centromeric plasmids with TRP1 marker, and with a doxycycline repressible promoter [Gari et al 1996]. <br />
# '''pEG202''', with a 2 ori, HIS3 marker and a constitutive promoter (PADH1). <br />
<br />
<br />
{| style="width:100%"<br />
|[[File:BsAs2012-plasmid-PEG202.jpg|400px]]<br />
|[[File:BsAs2012-plasmid-BPCM185.gif|340px]]<br />
|}<br />
<br />
The cloning we did was:<br />
<br />
{| class="wikitable"<br />
|<br />
! scope="row" style="background: #7ac5e8"|<partinfo>BBa_K792009</partinfo> (PoliHa)<br />
! scope="row" style="background: #7ac5e8"|<partinfo>BBa_K792010</partinfo> (TRPZipper2)<br />
! scope="row" style="background: #7ac5e8"|<partinfo>BBa_K792011</partinfo> (PoliHb)<br />
! scope="row" style="background: #7ac5e8"|<partinfo>BBa_K792012</partinfo> (PoliWb)<br />
|-<br />
! scope="row" style="background: #81BEF7"|pCM182 (TRPa)<br />
|<br />
! scope="row" style="background: #01DF01"|X<br />
|<br />
! scope="row" style="background: #01DF01"|X<br />
|-<br />
! scope="row" style="background: #81BEF7"|pCM185 (TRPb)<br />
|<br />
! scope="row" style="background: #01DF01"|X<br />
|<br />
! scope="row" style="background: #01DF01"|X<br />
|-<br />
! scope="row" style="background: #81BEF7"|pEG202 (HIS)<br />
! scope="row" style="background: #01DF01"|X<br />
|<br />
! scope="row" style="background: #01DF01"|X<br />
|<br />
|}<br />
<br />
<br />
==== Cloning protocol ====<br />
<br />
# Digestion of plasmids and TRP/HIS export device:<br />
##pCM182; pCM 185; BBa_K792010 y BBa_K792012 were digested with BamHI and Pst1 restriction enzymes.<br />
##pEG202; BBa_K792009 y BBa_K792011 were digested with BamHI and Not1.<br />
# Purification of digested vectors (pCM185; pCM182; pEG202) <br />
# Ligation of vectors and devices according the anterior table (T4 ligase protocol, overnight)<br />
# Transformation of E. Coli DH5a with the ligation products. Bacterias were plated on LB-agar with Ampicillin, and incubated over night at 37 °C.<br />
# Colonies were used for liquid cultures (LB + Ampicillin) and minipreps were made.<br />
# Constructions (vector + insert) were checked by digestion with restriction enzymes, and 1%-agarose gel (1kb and 100bp as markers).<br />
<br />
=== Yeast strains ===<br />
<br />
Once obtained the desired constructions, we transformed yeast strains:<br />
# TCY3081: W303, bar1-, ura3::PAct1-YFP<br />
# TCY3128: W303, bar1-, leu2:: Pprm1-CFP 405<br />
<br />
<br />
<br />
=== Final transformed strains ===<br />
<br />
We got the following '''transformed strains''':<br />
<br />
{|<br />
|<br />
{| style="width:50%"<br />
| rowspan="3" | [[File:BsAs2012-icono-Cepa1.jpg | 200px]]<br />
! scope="row" style="background: #7ac5e8"| Name<br />
| '''YFP_TRPa_TRPZipper2'''<br />
|-<br />
! scope="row" style="background: #CEE3F6"|Strain<br />
|TCY3081 (YFP)<br />
|-<br />
! scope="row" style="background: #CEE3F6"|Plasmid<br />
|pCM182 (TRPa) + <partinfo>BBa_K792010</partinfo> (TRPZipper2)<br />
|}<br />
|<br />
{| style="width:50%"<br />
| rowspan="3" | [[File:BsAs2012-icono-Cepa2.jpg | 200px]]<br />
! scope="row" style="background: #7ac5e8"| Name<br />
| '''YFP_TRPa_PoliWb'''<br />
|-<br />
! scope="row" style="background: #CEE3F6"|Strain<br />
|TCY3081 (YFP)<br />
|-<br />
! scope="row" style="background: #CEE3F6"|Plasmid<br />
|pCM182 (TRPa) + <partinfo>BBa_K792012</partinfo> (PoliWb)<br />
|}<br />
|}<br />
<br />
{|<br />
|<br />
{| style="width:50%"<br />
| rowspan="3" | [[File:BsAs2012-icono-Cepa3.jpg | 200px]]<br />
! scope="row" style="background: #7ac5e8"| Name<br />
| '''YFP_TRPb_TRPZipper2'''<br />
|-<br />
! scope="row" style="background: #CEE3F6"|Strain<br />
|TCY3081 (YFP)<br />
|-<br />
! scope="row" style="background: #CEE3F6"|Plasmid<br />
|pCM185 (TRPb) + <partinfo>BBa_K792010</partinfo> (TRPZipper2)<br />
|}<br />
|<br />
{| style="width:50%"<br />
| rowspan="3" | [[File:BsAs2012-icono-Cepa4.jpg | 200px]]<br />
! scope="row" style="background: #7ac5e8"| Name<br />
| '''YFP_TRPb_PoliWb'''<br />
|-<br />
! scope="row" style="background: #CEE3F6"|Strain<br />
|TCY3081 (YFP)<br />
|-<br />
! scope="row" style="background: #CEE3F6"|Plasmid<br />
|pCM185 (TRPb) + <partinfo>BBa_K792012</partinfo> (PoliWb)<br />
|}<br />
|}<br />
<br />
{|<br />
|<br />
{| style="width:50%"<br />
| rowspan="3" | [[File:BsAs2012-icono-Cepa6.jpg | 200px]]<br />
! scope="row" style="background: #7ac5e8"| Name<br />
| '''CFP_HIS_PoliHa'''<br />
|-<br />
! scope="row" style="background: #CEE3F6"|Strain<br />
|TCY3128 (CFP)<br />
|-<br />
! scope="row" style="background: #CEE3F6"|Plasmid<br />
|pEG202 (HIS) + <partinfo>BBa_K792009</partinfo> (PoliHa)<br />
|}<br />
|<br />
{| style="width:50%"<br />
| rowspan="3" | [[File:BsAs2012-icono-Cepa5.jpg | 200px]]<br />
! scope="row" style="background: #7ac5e8"| Name<br />
| '''CFP_HIS_PoliHb'''<br />
|-<br />
! scope="row" style="background: #CEE3F6"|Strain<br />
|TCY3128 (CFP)<br />
|-<br />
! scope="row" style="background: #CEE3F6"|Plasmid<br />
|pEG202 (HIS) + <partinfo>BBa_K792011</partinfo> (PoliHb)<br />
|}<br />
|}<br />
<br />
<br />
<br />
== Secretion Rate of Trp as a function of culture growth ==<br />
<br />
The first step was to actually check if the construct works: do the transformed yeast strains - with the tryptophan devices- actually secrete tryptophan into the medium?<br />
<br />
To test this we used the following strains:<br />
<br />
{|<br />
|<br />
{|<br />
|-<br />
|[[File:BsAs2012-icono-Cepa3.jpg|200px]]<br />
|[[File:BsAs2012-icono-Cepa4.jpg|200px]]<br />
|[[File:BsAs2012-icono-YFP-185.jpg|200px]]<br />
|- align="center"<br />
|YFP_TRPb_TRPZipper2<br />
|YFP_TRPb_PolyWb<br />
|YFP_TRPb<br />
|}<br />
| rowspan="2" |<br />
{|<br />
| [[File:BsAs2012-icono-YFP.jpg | 200px]]<br />
|- align="center"<br />
|YFP<br />
|}<br />
<br />
|-<br />
|<br />
{|<br />
|-<br />
|[[File:BsAs2012-icono-Cepa1.jpg|200px]]<br />
|[[File:BsAs2012-icono-Cepa2.jpg|200px]]<br />
|[[File:BsAs2012-icono-YFP-182.jpg | 200px]]<br />
|- align="center"<br />
|YFP_TRPa_TRPZipper2<br />
|YFP_TRPa_PolyWb<br />
|YFP_TRPa<br />
|}<br />
<br />
|}<br />
<br />
<br />
===== Protocol =====<br />
<br />
*We started 5ml cultures with 3 replica until they reached exponential phase, overnight, using a -T medium.<br />
*Starting OD for the assay 0.1 (exponential phase). <br />
*We measured OD every hour until they reached an OD: 0.8 (5 hs approximately). <br />
*We measured the Trp signal for each culture medium using the spectrofluorometer.<br />
<br />
NOTE: The fluorescence meausurements taken for the Tryptophan in the medium, take into account both the aminoacid secreted by the device and the one diffused. <br />
<br />
Therefore the secretion rates calculated will be higher than the actual ones. We used an empty plasmid as control to study tryptophan diffusion.<br />
<br />
We used a simple model to measure the secretation rate for all the strains. Since the cultures are in exponential phase, we take<br />
<br />
[[File:BsAs2012-eqTrp1.jpg | 225px]]<br />
<br />
After a few calculations, we find that<br />
<br />
[[File:BsAs2012-eqTrp2.jpg | 225px]]<br />
<br />
===== Results =====<br />
<br />
Next we show the average OD for each strains used, needed to calculate the export rates of Trytophan.<br />
<br />
{|<br />
[[File:BsAs2012Odvstimea4.jpg | 250px]]<br />
|<br />
[[File:BsAs2012Odvstimeab.jpg | 250px]]<br />
|}<br />
Figure1. Cell density vs time for different strains in media that lack tryptophan or in synthetic defined medium (SC).<br />
<br />
We can see the cell groth for strain YFP transformed with different tryptophan devices or with empty plasmid, in media that lack tryptophan (-H) or in Synthetic Define medium (SC) containing tryptophan<br />
<br />
[[File:BsAs2012rate2.jpg| 450px]]<br />
<br />
Figure2. Tryptophan export rate for different tryptophan devices or with empty plasmids.<br />
<br />
We can see that YFP strain transformed with tryptophan export devices hace higher secretion rates, compared with the YFP yeast transformed with empty plasmid that should only diffuse tryptophan through the membrane. <br />
This negative control is about 10% of secretion rate found for the strains with devices and is the same order as the error associated.<br />
<br />
== Tryptophan secretion at increasing histidine concentrations ==<br />
<br />
We asked ourselves which was the dependance of tryptophan secretion on the amount of histidine in medium. We carried on this test in order to determine whether the secretion of tryptophan depends on the concentration of another aminoacid in the media, such as histidine and what would be the necessary amount of histidine in medium for the start of our system. <br />
<br />
In this experiment we used strains: <br />
<br />
<br />
{|<br />
|-<br />
|[[File:BsAs2012-icono-Cepa3.jpg|200px]]<br />
|[[File:BsAs2012-icono-Cepa4.jpg|200px]]<br />
|[[File:BsAs2012-icono-YFP-185.jpg|200px]]<br />
|- align="center"<br />
|YFP_TRPb_TRPZipper2<br />
|YFP_TRPb_PolyWb<br />
|YFP_TRPb (control)<br />
|}<br />
<br />
<br />
<br />
<br />
<br />
<br />
<br />
===== Protocol =====<br />
<br />
# Starters of each strain used were grown over night in -T media, at 30 °C in shaker.<br />
# After 12 hs, cells were pelleted and washed with -HT media.<br />
# Cultures with increasing concentrations of histidine (0X, 1X, 1/4X and 1/16X) were set at an initial OD: 0.1, for each strain with 2 replica.<br />
# We left the cultures in shaker at 30ºC for 5 hours. After that time, we measured the final OD reached by cultures with the use of a spectrophotometer and the amount of tryptophan present in medium with a spectrofluorometer.<br />
<br />
===== Results =====<br />
<br />
[[File: BsAs2012TrpvHis1.jpg|350px]]<br />
<br />
Figure 3. Final tryptophan concentration as a function of histidine concentration in the media.<br />
<br />
<br />
With this result we can infer that tryptophan export has no clear dependence on histidine concentration in the media. Therefore, supporting the assumption of the model.<br />
<br />
== Measurement of Trp in medium and Basal Production ==<br />
<br />
To check the efectiveness of our biobricks, we must first determine the ammount of tryptophan secreted by natural strains to the medium, so we can compare. With that end in mind, we designed a protocol for measurement of tryptophan in medium, based in its fluorescense at 350nm, when excited with 295nm light.<br />
As a previous step, we checked that none of the other aminoacids used in the medium interferes, by graphically comparing the spectres for uncomplemented medium and medium complemented with leucine, uracile and histidine, at an appropiate range.<br />
<br />
To determine Trp concentration, we must first have a way to transform our readings (intensity) to a more useful output, so we made a calibration curve, through serialized 1:2 dilutions of our medium, which Trp's concentration is 50mg/mL, until approximately constant intensity.<br />
<br />
The procedure to measure secretion rates will be growing the strain from a known OD in exponential growth phase in -T medium and plotting it's OD over time, spin-drying at time=t, retrieving the supernatant's Trp concentration and dividing it by the integral of OD vs. time between time=0 and time=t, so we get to a rate which will be proportional to the number of cells in the culture, which means we can actually compare between different strains. Since our medium is free from Trp, all of it should come from within the cells, and if the culture is growing at exponential rates, lysis should be negligible, so the only explanation would be cells exporting their own Trp.<br />
<br />
<br />
{| class="wikitable" border="1"<br />
|-<br />
|<!--column1-->[[File:Trp-bsas2012.png | 250px]]<br />
|-<br />
|Graph:Tryptophan calibration curve<br />
|}<br />
<br />
<br />
<br />
==== Results ====<br />
<br />
As can be seen from the graph the screening of the concentration of the Trp in medium describes an almost lineal function. Through this experiment we can be sure that we would be able to measure increase of Trp in medium as it is exported from the cells, within the biological range of export.<br />
The sensitivity of this method seems to be enough to detect concentrations as low as ~0.02mg/mL, and as high as 50mg/mL, maybe more. Since our medium is 50mg/mL, we assume that's the saturation point of the curve. If we get bigger intensities than the one corresponding to it, we will dilute the sample.<br />
<br />
Because of time constraints, we haven't been able to check the method with either our designed strains nor the non-exporting ones.</div>Lucho.Morohttp://2012.igem.org/Team:Buenos_Aires/Results/BBsTestingTeam:Buenos Aires/Results/BBsTesting2012-10-27T03:39:31Z<p>Lucho.Moro: /* Protocol */</p>
<hr />
<div>{{:Team:Buenos_Aires/Templates/menu}}<br />
= After the Jamboree! =<br />
<br />
<br />
DEVICE TESTING<br />
<br />
https://2012.igem.org/Team:Buenos_Aires/Results/DevicesTesting<br />
<br />
<br />
SYNECO TESTING<br />
<br />
https://2012.igem.org/Team:Buenos_Aires/Results/SynEcoTesting<br />
<br />
<br />
<br />
When we returned from the Latin America Jamboree we focused all our efforts on completing the neccesary transformations to test our devices and our project as a whole. <br />
<br />
We devided this task in three sections<br />
<br />
<br />
== Week 1&2 : Yeast expression vectors & Transformations ==<br />
<br />
In order to construct the yeast expression plasmids we choosed 3 vectors, 2 with a tryptophan marker and 1 with an histidine marker:<br />
# '''pCM182/5''', which are centromeric plasmids with TRP1 marker, and with a doxycycline repressible promoter [Gari et al 1996]. <br />
# '''pEG202''', with a 2 ori, HIS3 marker and a constitutive promoter (PADH1). <br />
<br />
<br />
{| style="width:100%"<br />
|[[File:BsAs2012-plasmid-PEG202.jpg|400px]]<br />
|[[File:BsAs2012-plasmid-BPCM185.gif|340px]]<br />
|}<br />
<br />
The cloning we did was:<br />
<br />
{| class="wikitable"<br />
|<br />
! scope="row" style="background: #7ac5e8"|<partinfo>BBa_K792009</partinfo> (PoliHa)<br />
! scope="row" style="background: #7ac5e8"|<partinfo>BBa_K792010</partinfo> (TRPZipper2)<br />
! scope="row" style="background: #7ac5e8"|<partinfo>BBa_K792011</partinfo> (PoliHb)<br />
! scope="row" style="background: #7ac5e8"|<partinfo>BBa_K792012</partinfo> (PoliWb)<br />
|-<br />
! scope="row" style="background: #81BEF7"|pCM182 (TRPa)<br />
|<br />
! scope="row" style="background: #01DF01"|X<br />
|<br />
! scope="row" style="background: #01DF01"|X<br />
|-<br />
! scope="row" style="background: #81BEF7"|pCM185 (TRPb)<br />
|<br />
! scope="row" style="background: #01DF01"|X<br />
|<br />
! scope="row" style="background: #01DF01"|X<br />
|-<br />
! scope="row" style="background: #81BEF7"|pEG202 (HIS)<br />
! scope="row" style="background: #01DF01"|X<br />
|<br />
! scope="row" style="background: #01DF01"|X<br />
|<br />
|}<br />
<br />
<br />
==== Protocol ====<br />
<br />
# Digestion of plasmids and TRP/HIS export device:<br />
##pCM182; pCM 185; BBa_K792010 y BBa_K792012 were digested with BamHI and Pst1 restriction enzymes.<br />
##pEG202; BBa_K792009 y BBa_K792011 were digested with BamHI and Not1.<br />
# Purification of digested vectors (pCM185; pCM182; pEG202) <br />
# Ligation of vectors and devices according the anterior table (T4 ligase protocol, overnight)<br />
# Transformation of E. Coli DH5a with the ligation products. Bacterias were plated on LB-agar with Ampicillin, and incubated over night at 37 °C.<br />
# Colonies were used for liquid cultures (LB + Ampicillin) and minipreps were made.<br />
# Constructions (vector + insert) were checked by digestion with restriction enzymes, and 1%-agarose gel (1kb and 100bp as markers).<br />
<br />
<br />
Once obtained the desired constructions, we transformed yeast strains:<br />
# TCY3081: W303, bar1-, ura3::PAct1-YFP<br />
# TCY3128: W303, bar1-, leu2:: Pprm1-CFP 405<br />
<br />
<br />
<br />
We got the following '''transformed strains''':<br />
<br />
{|<br />
|<br />
{| style="width:50%"<br />
| rowspan="3" | [[File:BsAs2012-icono-Cepa1.jpg | 200px]]<br />
! scope="row" style="background: #7ac5e8"| Name<br />
| '''YFP_TRPa_TRPZipper2'''<br />
|-<br />
! scope="row" style="background: #CEE3F6"|Strain<br />
|TCY3081 (YFP)<br />
|-<br />
! scope="row" style="background: #CEE3F6"|Plasmid<br />
|pCM182 (TRPa) + <partinfo>BBa_K792010</partinfo> (TRPZipper2)<br />
|}<br />
|<br />
{| style="width:50%"<br />
| rowspan="3" | [[File:BsAs2012-icono-Cepa2.jpg | 200px]]<br />
! scope="row" style="background: #7ac5e8"| Name<br />
| '''YFP_TRPa_PoliWb'''<br />
|-<br />
! scope="row" style="background: #CEE3F6"|Strain<br />
|TCY3081 (YFP)<br />
|-<br />
! scope="row" style="background: #CEE3F6"|Plasmid<br />
|pCM182 (TRPa) + <partinfo>BBa_K792012</partinfo> (PoliWb)<br />
|}<br />
|}<br />
<br />
{|<br />
|<br />
{| style="width:50%"<br />
| rowspan="3" | [[File:BsAs2012-icono-Cepa3.jpg | 200px]]<br />
! scope="row" style="background: #7ac5e8"| Name<br />
| '''YFP_TRPb_TRPZipper2'''<br />
|-<br />
! scope="row" style="background: #CEE3F6"|Strain<br />
|TCY3081 (YFP)<br />
|-<br />
! scope="row" style="background: #CEE3F6"|Plasmid<br />
|pCM185 (TRPb) + <partinfo>BBa_K792010</partinfo> (TRPZipper2)<br />
|}<br />
|<br />
{| style="width:50%"<br />
| rowspan="3" | [[File:BsAs2012-icono-Cepa4.jpg | 200px]]<br />
! scope="row" style="background: #7ac5e8"| Name<br />
| '''YFP_TRPb_PoliWb'''<br />
|-<br />
! scope="row" style="background: #CEE3F6"|Strain<br />
|TCY3081 (YFP)<br />
|-<br />
! scope="row" style="background: #CEE3F6"|Plasmid<br />
|pCM185 (TRPb) + <partinfo>BBa_K792012</partinfo> (PoliWb)<br />
|}<br />
|}<br />
<br />
{|<br />
|<br />
{| style="width:50%"<br />
| rowspan="3" | [[File:BsAs2012-icono-Cepa6.jpg | 200px]]<br />
! scope="row" style="background: #7ac5e8"| Name<br />
| '''CFP_HIS_PoliHa'''<br />
|-<br />
! scope="row" style="background: #CEE3F6"|Strain<br />
|TCY3128 (CFP)<br />
|-<br />
! scope="row" style="background: #CEE3F6"|Plasmid<br />
|pEG202 (HIS) + <partinfo>BBa_K792009</partinfo> (PoliHa)<br />
|}<br />
|<br />
{| style="width:50%"<br />
| rowspan="3" | [[File:BsAs2012-icono-Cepa5.jpg | 200px]]<br />
! scope="row" style="background: #7ac5e8"| Name<br />
| '''CFP_HIS_PoliHb'''<br />
|-<br />
! scope="row" style="background: #CEE3F6"|Strain<br />
|TCY3128 (CFP)<br />
|-<br />
! scope="row" style="background: #CEE3F6"|Plasmid<br />
|pEG202 (HIS) + <partinfo>BBa_K792011</partinfo> (PoliHb)<br />
|}<br />
|}<br />
<br />
== Week 3: Synthetic Ecology Characterization ==<br />
<br />
Our task is to characterize our system including the devices functioning.<br />
We want specifically to: <br />
<br />
#Quantify the export of Trp as proof that the devices work<br />
#Determine experimentally some of the parameters that we used in the model<br />
#Characterize the growth of the transformed strains in coculture<br />
<br />
In order to characterize the devices that we used to transform our strains we came up with the series of assays that we describe below.<br />
<br />
<br />
<br />
=== Devices characterization ===<br />
<br />
==== Secretion Rate of Trp as a function of culture growth ====<br />
<br />
The first step was to actually check if the construct works: do the transformed yeast strains - with the tryptophan devices- actually secrete tryptophan into the medium?<br />
<br />
To test this we used the following strains:<br />
<br />
{|<br />
|<br />
{|<br />
|-<br />
|[[File:BsAs2012-icono-Cepa3.jpg|200px]]<br />
|[[File:BsAs2012-icono-Cepa4.jpg|200px]]<br />
|[[File:BsAs2012-icono-YFP-185.jpg|200px]]<br />
|- align="center"<br />
|YFP_TRPb_TRPZipper2<br />
|YFP_TRPb_PolyWb<br />
|YFP_TRPb<br />
|}<br />
| rowspan="2" |<br />
{|<br />
| [[File:BsAs2012-icono-YFP.jpg | 200px]]<br />
|- align="center"<br />
|YFP<br />
|}<br />
<br />
|-<br />
|<br />
{|<br />
|-<br />
|[[File:BsAs2012-icono-Cepa1.jpg|200px]]<br />
|[[File:BsAs2012-icono-Cepa2.jpg|200px]]<br />
|[[File:BsAs2012-icono-YFP-182.jpg | 200px]]<br />
|- align="center"<br />
|YFP_TRPa_TRPZipper2<br />
|YFP_TRPa_PolyWb<br />
|YFP_TRPa<br />
|}<br />
<br />
|}<br />
<br />
<br />
===== Protocol =====<br />
<br />
*We started 5ml cultures with 3 replica until they reached exponential phase, overnight, using a -T medium.<br />
*Starting OD for the assay 0.1 (exponential phase). <br />
*We measured OD every hour until they reached an OD: 0.8 (5 hs approximately). <br />
*We measured the Trp signal for each culture medium using the spectrofluorometer.<br />
<br />
NOTE: The fluorescence meausurements taken for the Tryptophan in the medium, take into account both the aminoacid secreted by the device and the one diffused. <br />
<br />
Therefore the secretion rates calculated will be higher than the actual ones. We used an empty plasmid as control to study tryptophan diffusion.<br />
<br />
We used a simple model to measure the secretion rate for all the strains. Since the cultures are in exponential phase, we take<br />
<br />
[[File:BsAs2012-eqTrp1.jpg | 225px]]<br />
<br />
After a few calculations, we find that<br />
<br />
[[File:BsAs2012-eqTrp2.jpg | 225px]]<br />
<br />
===== Results =====<br />
<br />
Next we show the average OD for each strains used, needed to calculate the secretation rates of Trytophan.<br />
<br />
{|<br />
[[File:BsAs2012Odvstimea4.jpg | 350px]]<br />
|<br />
[[File:BsAs2012Odvstimeab.jpg | 350px]]<br />
|}<br />
Figure X. check<br />
<br />
[[File:BsAs2012rate2.jpg| 350px]]<br />
<br />
Figure X. check<br />
<br />
==== Tryptophan secretion at increasing histidine concentrations ====<br />
<br />
We asked ourselves which was the dependance of tryptophan secretion on the amount of histidine in medium. We carried on this test in order to determine whether the secretion of tryptophan depends on the concentration of another aminoacid in the media, such as histidine and what would be the necessary amount of histidine in medium for the start of our system. <br />
<br />
In this experiment we used strains: <br />
<br />
<br />
{|<br />
|-<br />
|[[File:BsAs2012-icono-Cepa3.jpg|200px]]<br />
|[[File:BsAs2012-icono-Cepa4.jpg|200px]]<br />
|[[File:BsAs2012-icono-YFP-185.jpg|200px]]<br />
|- align="center"<br />
|YFP_TRPb_TRPZipper2<br />
|YFP_TRPb_PolyWb<br />
|YFP_TRPb (control)<br />
|}<br />
<br />
<br />
<br />
<br />
<br />
<br />
<br />
===== Protocol =====<br />
<br />
# Starters of each strain used were grown over night in -T media, at 30 °C in shaker.<br />
# After 12 hs, cells were pelleted and washed with -HT media.<br />
# Cultures with increasing concentrations of histidine (0X, 1X, 1/4X and 1/16X) were set at an initial OD: 0.1, for each strain with 2 replica.<br />
# We left the cultures in shaker at 30ºC for 5 hours. After that time, we measured the final OD reached by cultures with the use of a spectrophotometer and the amount of tryptophan present in medium with a spectrofluorometer.<br />
<br />
===== Results =====<br />
<br />
[[File: BsAs2012TrpvHis.jpg|350px]]<br />
<br />
[[File: BsAs2012Trpratio.jpg|350px]]<br />
<br />
[[File: BsAs2012ratioTrpvHisbypromotor.jpg|350px]]<br />
<br />
=== Strain characterization ===<br />
==== Experimental determination of strains death rate====<br />
<br />
We set out to determine how long can auxotroph cells[link] survive in media that lacks both Trytophan and Histidine. These values are the '''death''' parameters for CFP and YFP strains used in our model[link]. These were taken as equal in the mathematical analysis for simplicity but now we would like to test whether this approximation is accurate.<br />
<br />
Given that our system most likely will present a lag phase until a certain amount of both AmioAcids is accumulated in the media, will the cells be viable until this occurs? This is a neccesary check of our '' system's feasability''.<br />
<br />
===== Protocol =====<br />
<br />
For this experiment we used<br />
{|<br />
|-<br />
|[[File:BsAs2012-icono-YFP.jpg|200px]]<br />
|[[File:BsAs2012-icono-CFP.jpg|200px]]<br />
|- align="center"<br />
|YFP Strain<br />
|CFP Strain<br />
|}<br />
<br />
*We set cultures of the two auxotroph strains without being transformed (YFP and CFP) in medium –HT at an initial OD of 0.01. <br />
*Each day we plated the same amount of µl of the culture and counted the number of colonies obtain in each plate. We set 3 replica of each strain.<br />
<br />
===== Result =====<br />
<br />
<br />
{| class="wikitable"<br />
! scope="row" style="background: #7ac5e8" |Strain<br />
! scope="row" style="background: #7ac5e8" |Replica<br />
! scope="row" style="background: #7ac5e8" |Monday<br />
! scope="row" style="background: #7ac5e8" |Tuesday<br />
! scope="row" style="background: #7ac5e8" |Wednesday<br />
|-<br />
|CFP<br />
|1<br />
|260<br />
|320<br />
|285<br />
|-<br />
|CFP<br />
|2<br />
|267<br />
|314<br />
|76<br />
|-<br />
|CFP<br />
|3<br />
|413<br />
|362<br />
|278<br />
|-<br />
|YFP<br />
|1<br />
|230<br />
|316<br />
|688<br />
|-<br />
|YFP<br />
|2<br />
|291<br />
|194<br />
|524<br />
|-<br />
|YFP<br />
|3<br />
|449<br />
|344<br />
|725<br />
|}<br />
<br />
'''Table:''' Number of colonies counted per plate.<br />
<br />
We expect to see a decrease in the number of colonies - because of cell death. We found that this was not the case, in the experiment's time lapse. However we observed that the size of the colonies was smaller everyday as can be seen in the following pictures.<br />
<br />
[[File:Bsas2012kdeathcells.png| 500px]]<br />
<br />
<br />
We can infer from this data that though they have not died, they may have enter into a '''...Alan state'''. In this way cells can survive for a period of time in media defficient in amino acid (at least, during the time course of our experiment), but grow slower. Probably this would require more time than 3 days to observe significative cell dying.<br />
<br />
<br />
{|<br />
|[[File:BsAs2012_celldeath.png | 100px]]<br />
|[[File:BsAs2012_celldeath2.png| 100px]]<br />
|<br />
|}<br />
FigureX.<br />
<br />
==== Growth dependence on the Trp and His concentration====<br />
<br />
An important thing in order to characterize the system is the dependence of the growth rate on the culture with the concentration of the crossfeeding aminoacids, tryptophane (Trp) and histidine (His).<br />
<br />
This would allow us to estimate one of the parameters used in our model (the EC50, which is the amount of aminoacid at which the culture reaches half of the maximum growth).<br />
<br />
<br />
===== Protocol=====<br />
<br />
1. For this experiment we would use strains YFP and CFP (non-transformed, without devices).<br />
We started cultures of 5 ml of initial OD: 0.025 for:<br />
<br />
{| class="wikitable"<br />
| Strain YFP at the following media [Trp] = 1x; 0.5x; 0.25x; 0.125x; 0.0625x, 0.03125x<br />
|-<br />
| Strain CFP at the following media [His] = 1x; 0.5x; 0.25x; 0.125x; 0.0625x, 0.03125x<br />
|-<br />
| Strain YFP at Synthetic Complete media<br />
|-<br />
| Strain CFP at Synthetic Complete media<br />
|}<br />
<br />
2. Cultures would be left overnight (12 hs) at 30°C with agitation and we measured OD reached with the use of spectrophotometer.<br />
<br />
===== Results ===== <br />
To be done with our strains soon.<br />
<br />
=== Coculture of transformed strains ===<br />
<br />
We did the first experiment to test whether our system works and how two transformed strains grow together.<br />
<br />
We designed an assay to do so; following the growth of these strains:<br />
<br />
{|<br />
|-<br />
|[[File:BsAs2012-icono-CFP-202.jpg|200px]]<br />
|[[File:BsAs2012-icono-YFP-185.jpg|200px]]<br />
|- align="center"<br />
|CFP_His<br />
|YFP_TRPb<br />
|}<br />
<br />
<br />
<br />
{|<br />
|<br />
{| class="wikitable"<br />
|+ Transformed cells coculture and controls.<br />
|! scope="row" align="center" style="background: #7ac5e8"| '''Treatment'''<br />
|! scope="row" align="center" style="background: #7ac5e8"|'''Strain A'''<br />
|! scope="row" align="center" style="background: #7ac5e8"| '''Strain B'''<br />
|-<br />
|1<br />
|YFP_TRPb_TRPZipper2<br />
|CFP_HIS_PolyHa<br />
|-<br />
|2<br />
|YFP_TRPb_TRPZipper2<br />
|CFP_HIS<br />
|-<br />
|3<br />
|YFP_TRPb_TRPZipper2<br />
|! scope="row" style="background: #CCCCCC"|<br />
|-<br />
|4<br />
|YFP_TRPb<br />
|CFP_HIS_PolyHa<br />
|-<br />
|5<br />
|! scope="row" style="background: #CCCCCC"|<br />
|CFP_HIS_PolyHa<br />
|-<br />
|6<br />
|YFP_TRPb<br />
|CFP_HIS<br />
|-<br />
|7<br />
|! scope="row" style="background: #CCCCCC"|<br />
|CFP_HIS<br />
|-<br />
|8<br />
|YFP_TRPb<br />
|! scope="row" style="background: #CCCCCC"|<br />
|}<br />
|}<br />
<br />
==== Protocol ====<br />
{|<br />
|<br />
# Starters of strains were grown over night at 30°C, according the scheme showed in the table<br />
# The next day cultures were sonicated briefly in low power, and OD was measured in order to check they were in exponential phase.<br />
# Cells were centrifugated and then washed with medium –HT.<br />
# We set the culture of strains at OD: 0.02 in 5 ml of medium –HT with 3 replica, according to Table 1. <br />
# At 0, 1, 2, 3, 4, 5, 6, 7, 8 and 22 hours we took samples of 20 µl.<br />
# Samples were placed in a 384 wells plate, with 20 µl of cyclohexamide 2x (final concentration 1x) in each of the wells.<br />
# We used epifluorescence microscope in order to determinate the strain proportion of each coculture. The density of the culture was calculated based on the cell density at in each wells.<br />
|<br />
{|class="wikitable"<br />
|! scope="row" align="center" style="background: #7ac5e8"|'''Strain'''<br />
|! scope="row" align="center" style="background: #7ac5e8"|'''Media'''<br />
|-<br />
|YFP_TRPb_TRPZipper2<br />
|'''-T'''<br />
|-<br />
|YFP_TRPb<br />
|'''-T'''<br />
|-<br />
|CFP_HIS_PolyHa<br />
|'''-H'''<br />
|-<br />
|CFP_HIS<br />
|'''-H'''<br />
|}<br />
|}<br />
<br />
{| class="wikitable" style="width:50%"<br />
| align="center" | [[File:Bsas2012-Wells.png|500px]]<br />
|-<br />
| 384 wells plate to be used for epifluorescence microscope.<br />
|}<br />
<br />
==== Results ====<br />
<br />
[[File:BsAs2012coculture1.jpg | 600px]]<br />
<br />
[[File:BsAs2012coculture2.jpg| 600px]]</div>Lucho.Morohttp://2012.igem.org/Team:Buenos_Aires/Results/StrainsTeam:Buenos Aires/Results/Strains2012-10-27T03:34:50Z<p>Lucho.Moro: /* Measurement of Trp in medium and Basal Production */</p>
<hr />
<div>{{:Team:Buenos_Aires/Templates/menu}}<br />
<br />
<br />
<br />
== Description of strains ==<br />
<br />
Through our experiments we worked with the following strains kindly provided by [http://www.ifibyne.fcen.uba.ar/new/temas-de-investigacion/laboratorio-de-fisiologia-y-biologia-molecular-lfbm/biologia-de-sistemas/dr-alejandro-colman-lerner/ Alejandro Colman-Lerner's] Lab: <br />
<br />
{|<br />
|<br />
{| class="wikitable"<br />
! scope="row" style="background: #7ac5e8" | Strain ID<br />
! scope="row" style="background: #7ac5e8" |Relevant Auxotrophies<br />
! scope="row" style="background: #7ac5e8" |Fluorescence<br />
! scope="row" style="background: #7ac5e8" |Used as<br />
|-<br />
|TCY 3043<br />
| style="text-align: center;" |(-H-T)<br />
|No fluorescence<br />
|Negative control<br />
|-<br />
|TCY 3190<br />
| style="text-align: center;" |(+H-T)<br />
|YFP + (Induced CFP)<br />
|For coculture<br />
|-<br />
|TCY 3265<br />
| style="text-align: center;" |(-H+T)<br />
|CFP<br />
|For coculture<br />
|-<br />
|TCY 3154<br />
| style="text-align: center;" |(+H+T)<br />
|CFP +(induced YFP)<br />
|Positive Control<br />
|}<br />
| rowspan="2" style="text-align: center;" |<br />
{| class="wikitable" border="1"<br />
|-<br />
|<!--column1-->[[File:Bsas2012-strains-figura1.jpg|300px]]<br />
|}<br />
|- valign="top"<br />
|<br />
<br />
In the table we can see Hystidine (H) and Tryptophane (T) auxotrophies per strain, type of fluorescence and description of most common utilization during the experiments.<br />
<br />
Nearly 15 other similar strains were evaluated and discarded due to several reasons (low screening potentiality; requirement of hormones for fluorescence induction; high reverting rate of auxotrophies, among others)<br />
|}<br />
<br />
== Measurement of strains fluorescence ==<br />
<br />
We measured Strains 3281 (YFP) and 3265 (CFP) and got a spectrum of each one prooving that these strains can be distinguished by their fluorescence in culture. <br />
<br />
'''Reference graph'''<br />
Image: YFP and CFP Emission and Absorption Spectra. Obtained from http://flowcyt.salk.edu/fluo.html<br />
<br />
{| style="width: 100%"<br />
| align="center" | [[File:Bsas2012-strains-Refefluro.png|300px]]<br />
|}<br />
<br />
<br />
'''Results'''<br />
<br />
{| style="width: 100%"<br />
| align="center" | [[File:Fluoro- igembsas2012. strains.png|650px]]<br />
|-<br />
|'''CFP Fluorescence Screening and YFP Fluorescence Screening'''<br />
|}<br />
<br />
When measuring YFP Strain 3281, we can see a clear peak around 530 while when measuring CFP Strain 3265, we can see a clear peak around 500, as expected.<br />
<br />
<br />
<br />
'''Discussion'''<br />
<br />
We were able to measure fluorescence in strains 3281 and 3265 using the spectrofluorometer. However, we considered it would not be precise enough for the purposes of measuring cocultures at different proportions. We also noticed a high background noise produced by dead yeast cells at high concentrations, which would make it possible to measure in this way only at a short range of OD while the culture is at exponential phase.<br />
<br />
== Screening of strain proportion ==<br />
<br />
A more precise way of measuring the proportion of the strains, is with a epifluorescence microscope.<br />
<br />
We mixed strains 3281 (expresses YFP) and 3265 (expresses CFP) in different proportions and analized the images obtained in the microscope, where we counted cells with different fluorescences. We also did a negative control with a non fluorescent strain (TCY 379). <br />
<br />
'''Description of Mixtures'''<br />
<br />
Mix 1: Negative Control Mix 2: 80% CFP; 20%YFP Mix 3: 60% CFP; 40%YFP<br />
<br />
Mix 4: 50% CFP; 50%YFP Mix 5: 40% CFP; 60%YFP Mix 6: 20% CFP; 80%YFP<br />
<br />
<br />
'''Results'''<br />
<br />
<br />
<br />
{| style="width: 100%"<br />
| align="center" | [[File:Montage-annotated.jpg|900px]]<br />
|-<br />
| style="text-align: center;" | Mixtures showing YFP and CFP fluorescence. <br />
|}<br />
<br />
<br />
<br />
As shown by images 1-6, cells showing different fluorescences can be count and distinguished from each other in a mixture of strains, and this could be used to measure strains proportion in a coculture. <br />
<br />
<br />
'''Counting of cells'''<br />
<br />
{| class="wikitable"<br />
|-<br />
|Fluorescence<br />
|Mix 1<br />
|Mix 2<br />
|Mix 3<br />
|Mix 4<br />
|Mix 5<br />
|Mix 6<br />
|-<br />
|YFP<br />
|0 <br />
|23 <br />
|67 <br />
|115 <br />
|135 <br />
|110<br />
|-<br />
|CFP<br />
|0* <br />
|235 <br />
|82 <br />
|107 <br />
|99 <br />
|78<br />
|}<br />
<br />
The table shows the number of cells counted by expression of fluorescence YFP and CFP in the different mixtures 1-6. I can be observed that the amount of cells is near the proportion stablished by OD measures when preparing the mixtures. This results confirms that epifluorescence measures are reliable and suitable for our research.<br />
<br />
== Auxotrophy confirmation ==<br />
<br />
<br />
Several times during the experiments we control and checked if the auxotrophies in the selected strain were functional by plating all of them in medium deficient in aminoacids (-H; -T; -H-T and control +H+T). <br />
We observed differential growth according to expected due to the description of each strain in point a)<br />
<br />
{| class="wikitable" <br />
|+ Auxotrophy check<br />
|-<br />
|[[File:Bsas2012-strains-figura3.jpg|100px]]<br />
|[[File:Bsas2012-strains-figura2.png|100px]]<br />
|[[File:Bsas2012-strains-figura4.png|100px]]<br />
|[[File:Bsas2012-strains-figura5.png|100px]]<br />
|-<br />
| style="text-align: center;" | Medium complete<br />
| style="text-align: center;" | Medium without H<br />
| style="text-align: center;" | Medium without T<br />
| style="text-align: center;" | Medium without H and T<br />
|}<br />
<br />
<br />
We observed all the strains grew in the SC plate (top left) and only 3154 (+H+T) grew in the -H-T plate (bottom right). In the -T plate (bottom left), only those strains able to synthesize T grew (3265 and 3154) and in the -H plate (top right) only those able to produce H grew (3190 and 3154), as expected. This means our strains work according to their description. We did this several times during the months to check for reversions or contaminations.<br />
<br />
== Coculture in liquid medium ==<br />
<br />
We used for these experiment TCY3190(H+T-) and TCY3265(H-T+)<br />
Positive control: TCY3154 (H+T+) and negative control TCY3043(H-T-)<br />
<br />
==== At different initial OD and proportions ====<br />
<br />
Cultures were set at different initial concentrations (0.25, 0.1 and 0.01) and proportions (1:1; 1:9; 9:1). After 24 hs, we measured OD with the use of a spectrophotometer (two replicas) and we calculated the mean OD and a Growth factor (as Mean OD en time 1 over Initial OD time 0). <br />
<br />
<br />
{| class="wikitable"<br />
|+ Coculture at different initial OD and proportions (Days 0 and 1)<br />
! scope="row" style="background: #7ac5e8" | Coculture Proportion (H+T-):(H-T+) <br />
! scope="row" style="background: #7ac5e8" |Initial OD(t=0) <br />
! scope="row" style="background: #7ac5e8"|OD1 (t=1) <br />
! scope="row" style="background: #7ac5e8"|OD2 (t=1) <br />
! scope="row" style="background: #7ac5e8"|dilution used for measure t=1 <br />
! scope="row" style="background: #7ac5e8"|Mean OD <br />
! scope="row" style="background: #7ac5e8"|Growth Factor<br />
|-<br />
|01:01 <br />
|0,25 <br />
|0,32 <br />
|0,314 <br />
|10 <br />
|3,17 <br />
|12,68<br />
|-<br />
|09:01 <br />
|0,25 <br />
|0,148 <br />
|0,144 <br />
|10 <br />
|1,46 <br />
|5,84<br />
|-<br />
|01:09 <br />
|0,25 <br />
|0,138 <br />
|0,189 <br />
|10 <br />
|1,635 <br />
|6,54<br />
|-<br />
|01:01 <br />
|0,1 <br />
|0,109 <br />
|0,169 <br />
|10 <br />
|1,39 <br />
|13,9<br />
|-<br />
|09:01 <br />
|0,1 <br />
|0,04 <br />
|0,045 <br />
|10 <br />
|0,425 <br />
|4,25<br />
|-<br />
|01:09 <br />
|0,1 <br />
|0,067 <br />
|0,053 <br />
|10 <br />
|0,6 <br />
|6<br />
|-<br />
|01:01 <br />
|0,01 <br />
|0,067 <br />
|0,061 <br />
|1 <br />
|0,064 <br />
|6,4<br />
|-<br />
|09:01 <br />
|0,01 <br />
|0,056 <br />
|0,05 <br />
|1 <br />
|0,053 <br />
|5,3<br />
|-<br />
|01:09 <br />
|0,01 <br />
|0,074 <br />
|0,073 <br />
|1 <br />
|0,0735 <br />
|7,35<br />
|-<br />
|} <br />
<br />
<br />
{|<br />
|-<br />
|<!--column1-->[[File:HIS-BSAS2012.png|400px]]<br />
|}<br />
<br />
<br />
As shown in graph and table there is a basal growth that does not depend on the initial OD or strain proportion, of a growth factor of 6 approximately.<br />
However we observed a much higher growth at the proportion 1:1 when the initial OD 0.25 and 0.1. Therefore we can assume that at these proportions there is a natural cooperation between the strains and that should be the level of growth that we would like to assess through our bioengineering. Besides we would like to be able in the future to tune the strains in order to be able to obtain in the proportions 9:1 and 1:9 similar results to those obtained in the 1:1, at our own will.<br />
<br />
==== At the same initial OD: 0.2, followed over time ====<br />
<br />
We set the same cultures and cocultures as in point i), but starting all of them at the same OD: 0.2 and we followed them over 2 days. At day 1 we took pictures of them and at day 2 we measured the final OD. <br />
<br />
{| align="center" <br />
|- valign="top"<br />
|<br />
{| class="wikitable"<br />
|+ Cultures set at initial OD: 0.2 and measured over time (Days 0 and 2)<br />
! scope="row" style="background: #7ac5e8"|Strain<br />
! scope="row" style="background: #7ac5e8"|Day 0 <br />
! scope="row" style="background: #7ac5e8"|Day 2<br />
|-<br />
|TCY 3190 (-H+T) <br />
|0,2<br />
|2,92<br />
|-<br />
|TCY 3265 (+H-T) <br />
|0,2<br />
|0,19<br />
|-<br />
|Coculture of strains (TCY 3190- TCY 3265) <br />
|0,2<br />
|2,76<br />
|-<br />
|Negative control (TCY 3043 / -H-T) <br />
|0,2<br />
|0,6<br />
|-<br />
|Positive Control (TCY 3154/ +H+T) <br />
|0,2<br />
|2,54<br />
|}<br />
<br />
<br />
{| class="wikitable" border="1"<br />
|-<br />
|<!--column1-->[[File:Bsas2012-strains-wednesday.png|500px]]<br />
|-<br />
|Picture: Day 1 after starting cultures, shows different OD reached by strains. <br />
|}<br />
<br />
We repeated this experiment 4 times with different modifications: increasing the amount of days for up to a week, measuring every 12 hs instead of every 24 hs and using different strains. However, bacterial contaminations and the high rate of revertants prevented us from getting to a valid results in those cases, whereas the experiment up to day 2 always worked correctly. This denotes that we should assess the problem of contamination (for example including ampicilin in the cultures) and revertant rate (revising the design of the experiment or looking for more stable strains) as the impossibility to go further than day 2 may put limitations to some applications of the Synthetic Community.<br />
|}<br />
<br />
== Coculture in Agar and Revertant mutation control ==<br />
<br />
<br />
Through this experiment we aim to quantify the rate of revertants of each strain, and to asses if cross-feeding between a lawn of cells of one strain and colonies from and other strain is posible. <br />
<br />
We used petri dishes with agar medium with (+) and without (-) Trp and His as shown in the following table.<br />
<br />
We started a culture of each strain in synthetic complete medium, measured its OD 24 hs after the culture initiated, replaced the synthetic complete medium for one lacking both H and T (to avoid residual growth) and plated ~10^6 cells (lawn) or ~10^2 cells (seed) as shown by the following table (we considered OD600=1 represents 3*10^7 cells). <br />
At the same time, 3 controls (one for each strain) were carried in YPD complete medium to check the viability of each strain separately, and to estimate the seed CFU (colony formin units) more precisely. <br />
<br />
{| class="wikitable"<br />
! scope="row" style="background: #7ac5e8"|Medium H<br />
! scope="row" style="background: #7ac5e8"|Medium T<br />
! scope="row" style="background: #7ac5e8"|Lawn (10^6 cells) <br />
! scope="row" style="background: #7ac5e8"|Seed (10^2 cells) <br />
! scope="row" style="background: #7ac5e8"|Description of experiment <br />
! scope="row" style="background: #7ac5e8"|Results after 3 days - Replica 1<br />
! scope="row" style="background: #7ac5e8"|Results after 3 days - Replica 2<br />
|-<br />
|(-) <br />
|(+) <br />
|(-) <br />
|Strain –H+T <br />
|Control of His revertants <br />
|7 <br />
|7<br />
|-<br />
|-<br />
|(+)<br />
|(-)<br />
|(-)<br />
|Strain +H-T <br />
|Control of Trp revertants <br />
|2 <br />
|7<br />
|-<br />
|(-)<br />
|(-)<br />
|Strain +H-T<br />
|Strain –H+T<br />
|Coculture; we expect to see natural cooperation<br />
|960<br />
|800<br />
|-<br />
|(-)<br />
|(-)<br />
|Strain –H+T<br />
|Strain +H-T<br />
|Coculture; we expect to see natural cooperation<br />
|500<br />
|712<br />
|-<br />
|(-)<br />
|(-)<br />
|(-)<br />
|Strain +H+T<br />
|Viability of yeasts in medium<br />
|171<br />
|(-)<br />
|}<br />
<br />
'''Table: Shows description of each plate content and results in number of colonies counted by plate at day 3. YPD control results plates are not shown in the table'''. <br />
<br />
{| class="wikitable" border="1"<br />
|-<br />
|<!--column1-->[[File:Bsas2012-strains-placas2.jpg|300px]]<br />
|<!--column1-->[[File:Bsas2012-strains-placas1.jpg|300px]]<br />
|-<br />
|Petri Dishes<br />
| With marks of the counting of colonies<br />
|}<br />
<br />
<br />
==== Results ====<br />
The viability of the strains was high as expected, as well as the viability of a control positive strain in the –H-T medium. <br />
As shown in the table, we have a low, but existent, number of revertants from both his and trp auxotrophy strains. This number should be taken into account when interpreting the results from coculture growth after several days, given that the rate of revertants in liquid medium is probably the same. <br />
<br />
Growth in coculture was puzzling, as it resulted in more colonies than the expected. If cooperation was effective, we expected to see as many colonies as "seed" cells, not more. Revertion of cells from the "lawn" doesn't explain the number of colonies either. Probably a combination of both these effects are taking place.<br />
<br />
== Measurement of Trp in medium ==<br />
<br />
To check the efectiveness of our biobricks, we must first determine the ammount of tryptophan secreted by natural strains to the medium, so we can compare. With that end in mind, we designed a protocol for measurement of tryptophan in medium, based in its fluorescense at 350nm, when excited with 295nm light.<br />
As a previous step, we checked that none of the other aminoacids used in the medium interferes, by graphically comparing the spectres for uncomplemented medium and medium complemented with leucine, uracile and histidine, at an appropiate range.<br />
<br />
To determine Trp concentration, we must first have a way to transform our readings (intensity) to a more useful output, so we made a calibration curve, through serialized 1:2 dilutions of our medium, which Trp's concentration is 20μg/ml, until approximately constant intensity.<br />
<br />
The procedure to measure secretion rates will be growing the strain from a known OD in exponential growth phase in -T medium and plotting it's OD over time, spin-drying at time=t, retrieving the supernatant's Trp concentration and dividing it by the integral of OD vs. time between time=0 and time=t, so we get to a rate which will be proportional to the number of cells in the culture, which means we can actually compare between different strains. Since our medium is free from Trp, all of it should come from within the cells, and if the culture is growing at exponential rates, lysis should be negligible, so the only explanation would be cells exporting and diffusing their own Trp.<br />
<br />
<br />
{| class="wikitable" border="1"<br />
|-<br />
|<!--column1-->[[File:Curva.png | 250px]]<br />
|-<br />
|Graph:Tryptophan calibration curve<br />
|}<br />
<br />
<br />
<br />
==== Results ====<br />
<br />
Through this experiment we can be sure that we would be able to measure increase of Trp in medium as it is exported from the cells, within the biological range of export.<br />
The sensitivity of this method seems to be enough to detect concentrations as low as ~0.01μg/ml, and as high as 20μg/ml, maybe more. Since our medium is 20μg/ml, we assume that's the saturation point of the curve. If we get bigger intensities than the one corresponding to it, we will dilute the sample.<br />
<br />
== Growth dependence on the Trp and His concentrations ==<br />
<br />
A important thing to characterize of the system is the dependence of the growth rate of the culture with the concentration of the crossfeeding aminoacids, tryptophane (Trp) and histidine (His). To do this we measured the final OD after an overnight growth in medium with different concentrations of Trp and His. <br />
<br />
We used strain ACL-379, that is auxotroph for both Trp and His. <br />
We prepared serial dilutions of SC medium in –T and –H medium, therefore creating two curves: one with decreasing concentrations of Trp and the other with decreasing concentrations of His. <br />
We then inoculated equal amounts of ACL-379 in each tube and incubated them overnight at 30°C with agitation. We took a picture of each tube and measured the OD600 reached by each culture.<br />
<br />
{| class="wikitable"<br />
|+Growth of ACL-379 as a function of Trp and His concentration<br />
! scope="row" style="background: #7ac5e8"|Medium<br />
! scope="row" style="background: #7ac5e8"|OD Replica 1<br />
! scope="row" style="background: #7ac5e8"|OD Replica 2<br />
|-<br />
|SC (no cells)<br />
|0,001<br />
|(-0,0036)<br />
|-<br />
| -T<br />
|(-0,003)<br />
|(-0,019)<br />
|-<br />
| Trp/2<br />
|2.56 <br />
|2.17<br />
|-<br />
| Trp/4<br />
|3.01 <br />
|3.11<br />
|-<br />
|Trp/8<br />
|1.54 <br />
|1.55<br />
|-<br />
|Trp/16<br />
|0.393 <br />
|0.409<br />
|-<br />
|Trp/32<br />
|0.013 <br />
|0.003<br />
|-<br />
| -H<br />
|(-0,008) <br />
|(-0,012)<br />
|-<br />
| His/2 <br />
|3.68 <br />
|3.84<br />
|-<br />
| His/4<br />
|2.07 <br />
|2.00<br />
|-<br />
|His/8<br />
|1.17 <br />
|0.97<br />
|-<br />
|His/16 <br />
|0.47 <br />
|0.432<br />
|-<br />
|His/32 <br />
|0.238 <br />
|0.257<br />
|-<br />
|SC (w/cells) <br />
|4.88 <br />
|4.91<br />
|} <br />
<br />
==== Results ====<br />
As expected the growth has a sigmoidal relationship with the concentration of Trp and His, when plotted in semilogarithmic scale. We call EC50 the effective concentration of each aminoacid at which the culture reaches 50% of the maximal growth. We considered these values as proxies of the Khis and Ktrp parameters of the [[Team:Buenos_Aires/Project/Model#Parameter_selection|mathematical model]], which can be used to estimate the secretion rate of each aminoacid needed to get effective crossfeeding. <br />
<br />
These results can also be observed by comparison of images that show the tubes at different OD. <br />
<br />
{| class="wikitable" border="1"<br />
|-<br />
|<!--column1-->[[File:Bsas2012-strains-alan1.png|250px]]<br />
|<!--column2-->[[File:Bsas2012-strains-ultima.jpg|250px]] <br />
|-<br />
|Images from HLU series<br />
|Images from TLU series<br />
|}<br />
<br />
Notes: <br />
SC: Synthetic complete medium with all the aminoacids. It was used as a blank for the spectrofluorometer.<br />
<br />
HTLU is the culture in the medium with all the required aminoacids.<br />
<br />
<br />
== Experimental determination of strains death rate==<br />
<br />
We set out to determine how long can auxotroph cells[link] survive in media that lacks both Trytophan and Histidine. These values are the '''death''' parameters for CFP and YFP strains used in our model[link]. These were taken as equal in the mathematical analysis for simplicity but now we would like to test whether this approximation is accurate.<br />
<br />
Given that our system most likely will present a lag phase until a certain amount of both AmioAcids is accumulated in the media, will the cells be viable until this occurs? This is a neccesary check of our '' system's feasability''.<br />
<br />
===== Protocol =====<br />
<br />
For this experiment we used<br />
{|<br />
|-<br />
|[[File:BsAs2012-icono-YFP.jpg|200px]]<br />
|[[File:BsAs2012-icono-CFP.jpg|200px]]<br />
|- align="center"<br />
|YFP Strain<br />
|CFP Strain<br />
|}<br />
<br />
*We set cultures of the two auxotroph strains without being transformed (YFP and CFP) in medium –HT at an initial OD of 0.01. <br />
*Each day we plated the same amount of µl of the culture and counted the number of colonies obtain in each plate. We set 3 replica of each strain.<br />
<br />
===== Result =====<br />
<br />
<br />
{| class="wikitable"<br />
! scope="row" style="background: #7ac5e8" |Strain<br />
! scope="row" style="background: #7ac5e8" |Replica<br />
! scope="row" style="background: #7ac5e8" |Monday<br />
! scope="row" style="background: #7ac5e8" |Tuesday<br />
! scope="row" style="background: #7ac5e8" |Wednesday<br />
|-<br />
|CFP<br />
|1<br />
|260<br />
|320<br />
|285<br />
|-<br />
|CFP<br />
|2<br />
|267<br />
|314<br />
|76<br />
|-<br />
|CFP<br />
|3<br />
|413<br />
|362<br />
|278<br />
|-<br />
|YFP<br />
|1<br />
|230<br />
|316<br />
|688<br />
|-<br />
|YFP<br />
|2<br />
|291<br />
|194<br />
|524<br />
|-<br />
|YFP<br />
|3<br />
|449<br />
|344<br />
|725<br />
|}<br />
<br />
'''Table:''' Number of colonies counted per plate.<br />
<br />
We expect to see a decrease in the number of colonies - because of cell death. We found that this was not the case, in the experiment's time lapse. However we observed that the size of the colonies was smaller everyday as can be seen in the following pictures.<br />
<br />
[[File:Bsas2012kdeathcells.png| 500px]]<br />
<br />
<br />
We can infer from this data that though they have not died, they may have enter into a '''...Alan state'''. In this way cells can survive for a period of time in media defficient in amino acid (at least, during the time course of our experiment), but grow slower. Probably this would require more time than 3 days to observe significative cell dying.<br />
<br />
<br />
{|<br />
|[[File:BsAs2012_celldeath.png | 100px]]<br />
|[[File:BsAs2012_celldeath2.png| 100px]]<br />
|<br />
|}<br />
FigureX.</div>Lucho.Morohttp://2012.igem.org/Team:Buenos_Aires/Results/StrainsTeam:Buenos Aires/Results/Strains2012-10-27T03:34:27Z<p>Lucho.Moro: /* Measurement of Trp in medium and Basal Production */</p>
<hr />
<div>{{:Team:Buenos_Aires/Templates/menu}}<br />
<br />
<br />
<br />
== Description of strains ==<br />
<br />
Through our experiments we worked with the following strains kindly provided by [http://www.ifibyne.fcen.uba.ar/new/temas-de-investigacion/laboratorio-de-fisiologia-y-biologia-molecular-lfbm/biologia-de-sistemas/dr-alejandro-colman-lerner/ Alejandro Colman-Lerner's] Lab: <br />
<br />
{|<br />
|<br />
{| class="wikitable"<br />
! scope="row" style="background: #7ac5e8" | Strain ID<br />
! scope="row" style="background: #7ac5e8" |Relevant Auxotrophies<br />
! scope="row" style="background: #7ac5e8" |Fluorescence<br />
! scope="row" style="background: #7ac5e8" |Used as<br />
|-<br />
|TCY 3043<br />
| style="text-align: center;" |(-H-T)<br />
|No fluorescence<br />
|Negative control<br />
|-<br />
|TCY 3190<br />
| style="text-align: center;" |(+H-T)<br />
|YFP + (Induced CFP)<br />
|For coculture<br />
|-<br />
|TCY 3265<br />
| style="text-align: center;" |(-H+T)<br />
|CFP<br />
|For coculture<br />
|-<br />
|TCY 3154<br />
| style="text-align: center;" |(+H+T)<br />
|CFP +(induced YFP)<br />
|Positive Control<br />
|}<br />
| rowspan="2" style="text-align: center;" |<br />
{| class="wikitable" border="1"<br />
|-<br />
|<!--column1-->[[File:Bsas2012-strains-figura1.jpg|300px]]<br />
|}<br />
|- valign="top"<br />
|<br />
<br />
In the table we can see Hystidine (H) and Tryptophane (T) auxotrophies per strain, type of fluorescence and description of most common utilization during the experiments.<br />
<br />
Nearly 15 other similar strains were evaluated and discarded due to several reasons (low screening potentiality; requirement of hormones for fluorescence induction; high reverting rate of auxotrophies, among others)<br />
|}<br />
<br />
== Measurement of strains fluorescence ==<br />
<br />
We measured Strains 3281 (YFP) and 3265 (CFP) and got a spectrum of each one prooving that these strains can be distinguished by their fluorescence in culture. <br />
<br />
'''Reference graph'''<br />
Image: YFP and CFP Emission and Absorption Spectra. Obtained from http://flowcyt.salk.edu/fluo.html<br />
<br />
{| style="width: 100%"<br />
| align="center" | [[File:Bsas2012-strains-Refefluro.png|300px]]<br />
|}<br />
<br />
<br />
'''Results'''<br />
<br />
{| style="width: 100%"<br />
| align="center" | [[File:Fluoro- igembsas2012. strains.png|650px]]<br />
|-<br />
|'''CFP Fluorescence Screening and YFP Fluorescence Screening'''<br />
|}<br />
<br />
When measuring YFP Strain 3281, we can see a clear peak around 530 while when measuring CFP Strain 3265, we can see a clear peak around 500, as expected.<br />
<br />
<br />
<br />
'''Discussion'''<br />
<br />
We were able to measure fluorescence in strains 3281 and 3265 using the spectrofluorometer. However, we considered it would not be precise enough for the purposes of measuring cocultures at different proportions. We also noticed a high background noise produced by dead yeast cells at high concentrations, which would make it possible to measure in this way only at a short range of OD while the culture is at exponential phase.<br />
<br />
== Screening of strain proportion ==<br />
<br />
A more precise way of measuring the proportion of the strains, is with a epifluorescence microscope.<br />
<br />
We mixed strains 3281 (expresses YFP) and 3265 (expresses CFP) in different proportions and analized the images obtained in the microscope, where we counted cells with different fluorescences. We also did a negative control with a non fluorescent strain (TCY 379). <br />
<br />
'''Description of Mixtures'''<br />
<br />
Mix 1: Negative Control Mix 2: 80% CFP; 20%YFP Mix 3: 60% CFP; 40%YFP<br />
<br />
Mix 4: 50% CFP; 50%YFP Mix 5: 40% CFP; 60%YFP Mix 6: 20% CFP; 80%YFP<br />
<br />
<br />
'''Results'''<br />
<br />
<br />
<br />
{| style="width: 100%"<br />
| align="center" | [[File:Montage-annotated.jpg|900px]]<br />
|-<br />
| style="text-align: center;" | Mixtures showing YFP and CFP fluorescence. <br />
|}<br />
<br />
<br />
<br />
As shown by images 1-6, cells showing different fluorescences can be count and distinguished from each other in a mixture of strains, and this could be used to measure strains proportion in a coculture. <br />
<br />
<br />
'''Counting of cells'''<br />
<br />
{| class="wikitable"<br />
|-<br />
|Fluorescence<br />
|Mix 1<br />
|Mix 2<br />
|Mix 3<br />
|Mix 4<br />
|Mix 5<br />
|Mix 6<br />
|-<br />
|YFP<br />
|0 <br />
|23 <br />
|67 <br />
|115 <br />
|135 <br />
|110<br />
|-<br />
|CFP<br />
|0* <br />
|235 <br />
|82 <br />
|107 <br />
|99 <br />
|78<br />
|}<br />
<br />
The table shows the number of cells counted by expression of fluorescence YFP and CFP in the different mixtures 1-6. I can be observed that the amount of cells is near the proportion stablished by OD measures when preparing the mixtures. This results confirms that epifluorescence measures are reliable and suitable for our research.<br />
<br />
== Auxotrophy confirmation ==<br />
<br />
<br />
Several times during the experiments we control and checked if the auxotrophies in the selected strain were functional by plating all of them in medium deficient in aminoacids (-H; -T; -H-T and control +H+T). <br />
We observed differential growth according to expected due to the description of each strain in point a)<br />
<br />
{| class="wikitable" <br />
|+ Auxotrophy check<br />
|-<br />
|[[File:Bsas2012-strains-figura3.jpg|100px]]<br />
|[[File:Bsas2012-strains-figura2.png|100px]]<br />
|[[File:Bsas2012-strains-figura4.png|100px]]<br />
|[[File:Bsas2012-strains-figura5.png|100px]]<br />
|-<br />
| style="text-align: center;" | Medium complete<br />
| style="text-align: center;" | Medium without H<br />
| style="text-align: center;" | Medium without T<br />
| style="text-align: center;" | Medium without H and T<br />
|}<br />
<br />
<br />
We observed all the strains grew in the SC plate (top left) and only 3154 (+H+T) grew in the -H-T plate (bottom right). In the -T plate (bottom left), only those strains able to synthesize T grew (3265 and 3154) and in the -H plate (top right) only those able to produce H grew (3190 and 3154), as expected. This means our strains work according to their description. We did this several times during the months to check for reversions or contaminations.<br />
<br />
== Coculture in liquid medium ==<br />
<br />
We used for these experiment TCY3190(H+T-) and TCY3265(H-T+)<br />
Positive control: TCY3154 (H+T+) and negative control TCY3043(H-T-)<br />
<br />
==== At different initial OD and proportions ====<br />
<br />
Cultures were set at different initial concentrations (0.25, 0.1 and 0.01) and proportions (1:1; 1:9; 9:1). After 24 hs, we measured OD with the use of a spectrophotometer (two replicas) and we calculated the mean OD and a Growth factor (as Mean OD en time 1 over Initial OD time 0). <br />
<br />
<br />
{| class="wikitable"<br />
|+ Coculture at different initial OD and proportions (Days 0 and 1)<br />
! scope="row" style="background: #7ac5e8" | Coculture Proportion (H+T-):(H-T+) <br />
! scope="row" style="background: #7ac5e8" |Initial OD(t=0) <br />
! scope="row" style="background: #7ac5e8"|OD1 (t=1) <br />
! scope="row" style="background: #7ac5e8"|OD2 (t=1) <br />
! scope="row" style="background: #7ac5e8"|dilution used for measure t=1 <br />
! scope="row" style="background: #7ac5e8"|Mean OD <br />
! scope="row" style="background: #7ac5e8"|Growth Factor<br />
|-<br />
|01:01 <br />
|0,25 <br />
|0,32 <br />
|0,314 <br />
|10 <br />
|3,17 <br />
|12,68<br />
|-<br />
|09:01 <br />
|0,25 <br />
|0,148 <br />
|0,144 <br />
|10 <br />
|1,46 <br />
|5,84<br />
|-<br />
|01:09 <br />
|0,25 <br />
|0,138 <br />
|0,189 <br />
|10 <br />
|1,635 <br />
|6,54<br />
|-<br />
|01:01 <br />
|0,1 <br />
|0,109 <br />
|0,169 <br />
|10 <br />
|1,39 <br />
|13,9<br />
|-<br />
|09:01 <br />
|0,1 <br />
|0,04 <br />
|0,045 <br />
|10 <br />
|0,425 <br />
|4,25<br />
|-<br />
|01:09 <br />
|0,1 <br />
|0,067 <br />
|0,053 <br />
|10 <br />
|0,6 <br />
|6<br />
|-<br />
|01:01 <br />
|0,01 <br />
|0,067 <br />
|0,061 <br />
|1 <br />
|0,064 <br />
|6,4<br />
|-<br />
|09:01 <br />
|0,01 <br />
|0,056 <br />
|0,05 <br />
|1 <br />
|0,053 <br />
|5,3<br />
|-<br />
|01:09 <br />
|0,01 <br />
|0,074 <br />
|0,073 <br />
|1 <br />
|0,0735 <br />
|7,35<br />
|-<br />
|} <br />
<br />
<br />
{|<br />
|-<br />
|<!--column1-->[[File:HIS-BSAS2012.png|400px]]<br />
|}<br />
<br />
<br />
As shown in graph and table there is a basal growth that does not depend on the initial OD or strain proportion, of a growth factor of 6 approximately.<br />
However we observed a much higher growth at the proportion 1:1 when the initial OD 0.25 and 0.1. Therefore we can assume that at these proportions there is a natural cooperation between the strains and that should be the level of growth that we would like to assess through our bioengineering. Besides we would like to be able in the future to tune the strains in order to be able to obtain in the proportions 9:1 and 1:9 similar results to those obtained in the 1:1, at our own will.<br />
<br />
==== At the same initial OD: 0.2, followed over time ====<br />
<br />
We set the same cultures and cocultures as in point i), but starting all of them at the same OD: 0.2 and we followed them over 2 days. At day 1 we took pictures of them and at day 2 we measured the final OD. <br />
<br />
{| align="center" <br />
|- valign="top"<br />
|<br />
{| class="wikitable"<br />
|+ Cultures set at initial OD: 0.2 and measured over time (Days 0 and 2)<br />
! scope="row" style="background: #7ac5e8"|Strain<br />
! scope="row" style="background: #7ac5e8"|Day 0 <br />
! scope="row" style="background: #7ac5e8"|Day 2<br />
|-<br />
|TCY 3190 (-H+T) <br />
|0,2<br />
|2,92<br />
|-<br />
|TCY 3265 (+H-T) <br />
|0,2<br />
|0,19<br />
|-<br />
|Coculture of strains (TCY 3190- TCY 3265) <br />
|0,2<br />
|2,76<br />
|-<br />
|Negative control (TCY 3043 / -H-T) <br />
|0,2<br />
|0,6<br />
|-<br />
|Positive Control (TCY 3154/ +H+T) <br />
|0,2<br />
|2,54<br />
|}<br />
<br />
<br />
{| class="wikitable" border="1"<br />
|-<br />
|<!--column1-->[[File:Bsas2012-strains-wednesday.png|500px]]<br />
|-<br />
|Picture: Day 1 after starting cultures, shows different OD reached by strains. <br />
|}<br />
<br />
We repeated this experiment 4 times with different modifications: increasing the amount of days for up to a week, measuring every 12 hs instead of every 24 hs and using different strains. However, bacterial contaminations and the high rate of revertants prevented us from getting to a valid results in those cases, whereas the experiment up to day 2 always worked correctly. This denotes that we should assess the problem of contamination (for example including ampicilin in the cultures) and revertant rate (revising the design of the experiment or looking for more stable strains) as the impossibility to go further than day 2 may put limitations to some applications of the Synthetic Community.<br />
|}<br />
<br />
== Coculture in Agar and Revertant mutation control ==<br />
<br />
<br />
Through this experiment we aim to quantify the rate of revertants of each strain, and to asses if cross-feeding between a lawn of cells of one strain and colonies from and other strain is posible. <br />
<br />
We used petri dishes with agar medium with (+) and without (-) Trp and His as shown in the following table.<br />
<br />
We started a culture of each strain in synthetic complete medium, measured its OD 24 hs after the culture initiated, replaced the synthetic complete medium for one lacking both H and T (to avoid residual growth) and plated ~10^6 cells (lawn) or ~10^2 cells (seed) as shown by the following table (we considered OD600=1 represents 3*10^7 cells). <br />
At the same time, 3 controls (one for each strain) were carried in YPD complete medium to check the viability of each strain separately, and to estimate the seed CFU (colony formin units) more precisely. <br />
<br />
{| class="wikitable"<br />
! scope="row" style="background: #7ac5e8"|Medium H<br />
! scope="row" style="background: #7ac5e8"|Medium T<br />
! scope="row" style="background: #7ac5e8"|Lawn (10^6 cells) <br />
! scope="row" style="background: #7ac5e8"|Seed (10^2 cells) <br />
! scope="row" style="background: #7ac5e8"|Description of experiment <br />
! scope="row" style="background: #7ac5e8"|Results after 3 days - Replica 1<br />
! scope="row" style="background: #7ac5e8"|Results after 3 days - Replica 2<br />
|-<br />
|(-) <br />
|(+) <br />
|(-) <br />
|Strain –H+T <br />
|Control of His revertants <br />
|7 <br />
|7<br />
|-<br />
|-<br />
|(+)<br />
|(-)<br />
|(-)<br />
|Strain +H-T <br />
|Control of Trp revertants <br />
|2 <br />
|7<br />
|-<br />
|(-)<br />
|(-)<br />
|Strain +H-T<br />
|Strain –H+T<br />
|Coculture; we expect to see natural cooperation<br />
|960<br />
|800<br />
|-<br />
|(-)<br />
|(-)<br />
|Strain –H+T<br />
|Strain +H-T<br />
|Coculture; we expect to see natural cooperation<br />
|500<br />
|712<br />
|-<br />
|(-)<br />
|(-)<br />
|(-)<br />
|Strain +H+T<br />
|Viability of yeasts in medium<br />
|171<br />
|(-)<br />
|}<br />
<br />
'''Table: Shows description of each plate content and results in number of colonies counted by plate at day 3. YPD control results plates are not shown in the table'''. <br />
<br />
{| class="wikitable" border="1"<br />
|-<br />
|<!--column1-->[[File:Bsas2012-strains-placas2.jpg|300px]]<br />
|<!--column1-->[[File:Bsas2012-strains-placas1.jpg|300px]]<br />
|-<br />
|Petri Dishes<br />
| With marks of the counting of colonies<br />
|}<br />
<br />
<br />
==== Results ====<br />
The viability of the strains was high as expected, as well as the viability of a control positive strain in the –H-T medium. <br />
As shown in the table, we have a low, but existent, number of revertants from both his and trp auxotrophy strains. This number should be taken into account when interpreting the results from coculture growth after several days, given that the rate of revertants in liquid medium is probably the same. <br />
<br />
Growth in coculture was puzzling, as it resulted in more colonies than the expected. If cooperation was effective, we expected to see as many colonies as "seed" cells, not more. Revertion of cells from the "lawn" doesn't explain the number of colonies either. Probably a combination of both these effects are taking place.<br />
<br />
== Measurement of Trp in medium and Basal Production ==<br />
<br />
To check the efectiveness of our biobricks, we must first determine the ammount of tryptophan secreted by natural strains to the medium, so we can compare. With that end in mind, we designed a protocol for measurement of tryptophan in medium, based in its fluorescense at 350nm, when excited with 295nm light.<br />
As a previous step, we checked that none of the other aminoacids used in the medium interferes, by graphically comparing the spectres for uncomplemented medium and medium complemented with leucine, uracile and histidine, at an appropiate range.<br />
<br />
To determine Trp concentration, we must first have a way to transform our readings (intensity) to a more useful output, so we made a calibration curve, through serialized 1:2 dilutions of our medium, which Trp's concentration is 20μg/ml, until approximately constant intensity.<br />
<br />
The procedure to measure secretion rates will be growing the strain from a known OD in exponential growth phase in -T medium and plotting it's OD over time, spin-drying at time=t, retrieving the supernatant's Trp concentration and dividing it by the integral of OD vs. time between time=0 and time=t, so we get to a rate which will be proportional to the number of cells in the culture, which means we can actually compare between different strains. Since our medium is free from Trp, all of it should come from within the cells, and if the culture is growing at exponential rates, lysis should be negligible, so the only explanation would be cells exporting and diffusing their own Trp.<br />
<br />
<br />
{| class="wikitable" border="1"<br />
|-<br />
|<!--column1-->[[File:Curva.png | 250px]]<br />
|-<br />
|Graph:Tryptophan calibration curve<br />
|}<br />
<br />
<br />
<br />
==== Results ====<br />
<br />
Through this experiment we can be sure that we would be able to measure increase of Trp in medium as it is exported from the cells, within the biological range of export.<br />
The sensitivity of this method seems to be enough to detect concentrations as low as ~0.01μg/ml, and as high as 20μg/ml, maybe more. Since our medium is 20μg/ml, we assume that's the saturation point of the curve. If we get bigger intensities than the one corresponding to it, we will dilute the sample.<br />
<br />
== Growth dependence on the Trp and His concentrations ==<br />
<br />
A important thing to characterize of the system is the dependence of the growth rate of the culture with the concentration of the crossfeeding aminoacids, tryptophane (Trp) and histidine (His). To do this we measured the final OD after an overnight growth in medium with different concentrations of Trp and His. <br />
<br />
We used strain ACL-379, that is auxotroph for both Trp and His. <br />
We prepared serial dilutions of SC medium in –T and –H medium, therefore creating two curves: one with decreasing concentrations of Trp and the other with decreasing concentrations of His. <br />
We then inoculated equal amounts of ACL-379 in each tube and incubated them overnight at 30°C with agitation. We took a picture of each tube and measured the OD600 reached by each culture.<br />
<br />
{| class="wikitable"<br />
|+Growth of ACL-379 as a function of Trp and His concentration<br />
! scope="row" style="background: #7ac5e8"|Medium<br />
! scope="row" style="background: #7ac5e8"|OD Replica 1<br />
! scope="row" style="background: #7ac5e8"|OD Replica 2<br />
|-<br />
|SC (no cells)<br />
|0,001<br />
|(-0,0036)<br />
|-<br />
| -T<br />
|(-0,003)<br />
|(-0,019)<br />
|-<br />
| Trp/2<br />
|2.56 <br />
|2.17<br />
|-<br />
| Trp/4<br />
|3.01 <br />
|3.11<br />
|-<br />
|Trp/8<br />
|1.54 <br />
|1.55<br />
|-<br />
|Trp/16<br />
|0.393 <br />
|0.409<br />
|-<br />
|Trp/32<br />
|0.013 <br />
|0.003<br />
|-<br />
| -H<br />
|(-0,008) <br />
|(-0,012)<br />
|-<br />
| His/2 <br />
|3.68 <br />
|3.84<br />
|-<br />
| His/4<br />
|2.07 <br />
|2.00<br />
|-<br />
|His/8<br />
|1.17 <br />
|0.97<br />
|-<br />
|His/16 <br />
|0.47 <br />
|0.432<br />
|-<br />
|His/32 <br />
|0.238 <br />
|0.257<br />
|-<br />
|SC (w/cells) <br />
|4.88 <br />
|4.91<br />
|} <br />
<br />
==== Results ====<br />
As expected the growth has a sigmoidal relationship with the concentration of Trp and His, when plotted in semilogarithmic scale. We call EC50 the effective concentration of each aminoacid at which the culture reaches 50% of the maximal growth. We considered these values as proxies of the Khis and Ktrp parameters of the [[Team:Buenos_Aires/Project/Model#Parameter_selection|mathematical model]], which can be used to estimate the secretion rate of each aminoacid needed to get effective crossfeeding. <br />
<br />
These results can also be observed by comparison of images that show the tubes at different OD. <br />
<br />
{| class="wikitable" border="1"<br />
|-<br />
|<!--column1-->[[File:Bsas2012-strains-alan1.png|250px]]<br />
|<!--column2-->[[File:Bsas2012-strains-ultima.jpg|250px]] <br />
|-<br />
|Images from HLU series<br />
|Images from TLU series<br />
|}<br />
<br />
Notes: <br />
SC: Synthetic complete medium with all the aminoacids. It was used as a blank for the spectrofluorometer.<br />
<br />
HTLU is the culture in the medium with all the required aminoacids.<br />
<br />
<br />
== Experimental determination of strains death rate==<br />
<br />
We set out to determine how long can auxotroph cells[link] survive in media that lacks both Trytophan and Histidine. These values are the '''death''' parameters for CFP and YFP strains used in our model[link]. These were taken as equal in the mathematical analysis for simplicity but now we would like to test whether this approximation is accurate.<br />
<br />
Given that our system most likely will present a lag phase until a certain amount of both AmioAcids is accumulated in the media, will the cells be viable until this occurs? This is a neccesary check of our '' system's feasability''.<br />
<br />
===== Protocol =====<br />
<br />
For this experiment we used<br />
{|<br />
|-<br />
|[[File:BsAs2012-icono-YFP.jpg|200px]]<br />
|[[File:BsAs2012-icono-CFP.jpg|200px]]<br />
|- align="center"<br />
|YFP Strain<br />
|CFP Strain<br />
|}<br />
<br />
*We set cultures of the two auxotroph strains without being transformed (YFP and CFP) in medium –HT at an initial OD of 0.01. <br />
*Each day we plated the same amount of µl of the culture and counted the number of colonies obtain in each plate. We set 3 replica of each strain.<br />
<br />
===== Result =====<br />
<br />
<br />
{| class="wikitable"<br />
! scope="row" style="background: #7ac5e8" |Strain<br />
! scope="row" style="background: #7ac5e8" |Replica<br />
! scope="row" style="background: #7ac5e8" |Monday<br />
! scope="row" style="background: #7ac5e8" |Tuesday<br />
! scope="row" style="background: #7ac5e8" |Wednesday<br />
|-<br />
|CFP<br />
|1<br />
|260<br />
|320<br />
|285<br />
|-<br />
|CFP<br />
|2<br />
|267<br />
|314<br />
|76<br />
|-<br />
|CFP<br />
|3<br />
|413<br />
|362<br />
|278<br />
|-<br />
|YFP<br />
|1<br />
|230<br />
|316<br />
|688<br />
|-<br />
|YFP<br />
|2<br />
|291<br />
|194<br />
|524<br />
|-<br />
|YFP<br />
|3<br />
|449<br />
|344<br />
|725<br />
|}<br />
<br />
'''Table:''' Number of colonies counted per plate.<br />
<br />
We expect to see a decrease in the number of colonies - because of cell death. We found that this was not the case, in the experiment's time lapse. However we observed that the size of the colonies was smaller everyday as can be seen in the following pictures.<br />
<br />
[[File:Bsas2012kdeathcells.png| 500px]]<br />
<br />
<br />
We can infer from this data that though they have not died, they may have enter into a '''...Alan state'''. In this way cells can survive for a period of time in media defficient in amino acid (at least, during the time course of our experiment), but grow slower. Probably this would require more time than 3 days to observe significative cell dying.<br />
<br />
<br />
{|<br />
|[[File:BsAs2012_celldeath.png | 100px]]<br />
|[[File:BsAs2012_celldeath2.png| 100px]]<br />
|<br />
|}<br />
FigureX.</div>Lucho.Morohttp://2012.igem.org/Team:Buenos_Aires/Results/StrainsTeam:Buenos Aires/Results/Strains2012-10-27T03:33:04Z<p>Lucho.Moro: /* Measurement of Trp in medium and Basal Production */</p>
<hr />
<div>{{:Team:Buenos_Aires/Templates/menu}}<br />
<br />
<br />
<br />
== Description of strains ==<br />
<br />
Through our experiments we worked with the following strains kindly provided by [http://www.ifibyne.fcen.uba.ar/new/temas-de-investigacion/laboratorio-de-fisiologia-y-biologia-molecular-lfbm/biologia-de-sistemas/dr-alejandro-colman-lerner/ Alejandro Colman-Lerner's] Lab: <br />
<br />
{|<br />
|<br />
{| class="wikitable"<br />
! scope="row" style="background: #7ac5e8" | Strain ID<br />
! scope="row" style="background: #7ac5e8" |Relevant Auxotrophies<br />
! scope="row" style="background: #7ac5e8" |Fluorescence<br />
! scope="row" style="background: #7ac5e8" |Used as<br />
|-<br />
|TCY 3043<br />
| style="text-align: center;" |(-H-T)<br />
|No fluorescence<br />
|Negative control<br />
|-<br />
|TCY 3190<br />
| style="text-align: center;" |(+H-T)<br />
|YFP + (Induced CFP)<br />
|For coculture<br />
|-<br />
|TCY 3265<br />
| style="text-align: center;" |(-H+T)<br />
|CFP<br />
|For coculture<br />
|-<br />
|TCY 3154<br />
| style="text-align: center;" |(+H+T)<br />
|CFP +(induced YFP)<br />
|Positive Control<br />
|}<br />
| rowspan="2" style="text-align: center;" |<br />
{| class="wikitable" border="1"<br />
|-<br />
|<!--column1-->[[File:Bsas2012-strains-figura1.jpg|300px]]<br />
|}<br />
|- valign="top"<br />
|<br />
<br />
In the table we can see Hystidine (H) and Tryptophane (T) auxotrophies per strain, type of fluorescence and description of most common utilization during the experiments.<br />
<br />
Nearly 15 other similar strains were evaluated and discarded due to several reasons (low screening potentiality; requirement of hormones for fluorescence induction; high reverting rate of auxotrophies, among others)<br />
|}<br />
<br />
== Measurement of strains fluorescence ==<br />
<br />
We measured Strains 3281 (YFP) and 3265 (CFP) and got a spectrum of each one prooving that these strains can be distinguished by their fluorescence in culture. <br />
<br />
'''Reference graph'''<br />
Image: YFP and CFP Emission and Absorption Spectra. Obtained from http://flowcyt.salk.edu/fluo.html<br />
<br />
{| style="width: 100%"<br />
| align="center" | [[File:Bsas2012-strains-Refefluro.png|300px]]<br />
|}<br />
<br />
<br />
'''Results'''<br />
<br />
{| style="width: 100%"<br />
| align="center" | [[File:Fluoro- igembsas2012. strains.png|650px]]<br />
|-<br />
|'''CFP Fluorescence Screening and YFP Fluorescence Screening'''<br />
|}<br />
<br />
When measuring YFP Strain 3281, we can see a clear peak around 530 while when measuring CFP Strain 3265, we can see a clear peak around 500, as expected.<br />
<br />
<br />
<br />
'''Discussion'''<br />
<br />
We were able to measure fluorescence in strains 3281 and 3265 using the spectrofluorometer. However, we considered it would not be precise enough for the purposes of measuring cocultures at different proportions. We also noticed a high background noise produced by dead yeast cells at high concentrations, which would make it possible to measure in this way only at a short range of OD while the culture is at exponential phase.<br />
<br />
== Screening of strain proportion ==<br />
<br />
A more precise way of measuring the proportion of the strains, is with a epifluorescence microscope.<br />
<br />
We mixed strains 3281 (expresses YFP) and 3265 (expresses CFP) in different proportions and analized the images obtained in the microscope, where we counted cells with different fluorescences. We also did a negative control with a non fluorescent strain (TCY 379). <br />
<br />
'''Description of Mixtures'''<br />
<br />
Mix 1: Negative Control Mix 2: 80% CFP; 20%YFP Mix 3: 60% CFP; 40%YFP<br />
<br />
Mix 4: 50% CFP; 50%YFP Mix 5: 40% CFP; 60%YFP Mix 6: 20% CFP; 80%YFP<br />
<br />
<br />
'''Results'''<br />
<br />
<br />
<br />
{| style="width: 100%"<br />
| align="center" | [[File:Montage-annotated.jpg|900px]]<br />
|-<br />
| style="text-align: center;" | Mixtures showing YFP and CFP fluorescence. <br />
|}<br />
<br />
<br />
<br />
As shown by images 1-6, cells showing different fluorescences can be count and distinguished from each other in a mixture of strains, and this could be used to measure strains proportion in a coculture. <br />
<br />
<br />
'''Counting of cells'''<br />
<br />
{| class="wikitable"<br />
|-<br />
|Fluorescence<br />
|Mix 1<br />
|Mix 2<br />
|Mix 3<br />
|Mix 4<br />
|Mix 5<br />
|Mix 6<br />
|-<br />
|YFP<br />
|0 <br />
|23 <br />
|67 <br />
|115 <br />
|135 <br />
|110<br />
|-<br />
|CFP<br />
|0* <br />
|235 <br />
|82 <br />
|107 <br />
|99 <br />
|78<br />
|}<br />
<br />
The table shows the number of cells counted by expression of fluorescence YFP and CFP in the different mixtures 1-6. I can be observed that the amount of cells is near the proportion stablished by OD measures when preparing the mixtures. This results confirms that epifluorescence measures are reliable and suitable for our research.<br />
<br />
== Auxotrophy confirmation ==<br />
<br />
<br />
Several times during the experiments we control and checked if the auxotrophies in the selected strain were functional by plating all of them in medium deficient in aminoacids (-H; -T; -H-T and control +H+T). <br />
We observed differential growth according to expected due to the description of each strain in point a)<br />
<br />
{| class="wikitable" <br />
|+ Auxotrophy check<br />
|-<br />
|[[File:Bsas2012-strains-figura3.jpg|100px]]<br />
|[[File:Bsas2012-strains-figura2.png|100px]]<br />
|[[File:Bsas2012-strains-figura4.png|100px]]<br />
|[[File:Bsas2012-strains-figura5.png|100px]]<br />
|-<br />
| style="text-align: center;" | Medium complete<br />
| style="text-align: center;" | Medium without H<br />
| style="text-align: center;" | Medium without T<br />
| style="text-align: center;" | Medium without H and T<br />
|}<br />
<br />
<br />
We observed all the strains grew in the SC plate (top left) and only 3154 (+H+T) grew in the -H-T plate (bottom right). In the -T plate (bottom left), only those strains able to synthesize T grew (3265 and 3154) and in the -H plate (top right) only those able to produce H grew (3190 and 3154), as expected. This means our strains work according to their description. We did this several times during the months to check for reversions or contaminations.<br />
<br />
== Coculture in liquid medium ==<br />
<br />
We used for these experiment TCY3190(H+T-) and TCY3265(H-T+)<br />
Positive control: TCY3154 (H+T+) and negative control TCY3043(H-T-)<br />
<br />
==== At different initial OD and proportions ====<br />
<br />
Cultures were set at different initial concentrations (0.25, 0.1 and 0.01) and proportions (1:1; 1:9; 9:1). After 24 hs, we measured OD with the use of a spectrophotometer (two replicas) and we calculated the mean OD and a Growth factor (as Mean OD en time 1 over Initial OD time 0). <br />
<br />
<br />
{| class="wikitable"<br />
|+ Coculture at different initial OD and proportions (Days 0 and 1)<br />
! scope="row" style="background: #7ac5e8" | Coculture Proportion (H+T-):(H-T+) <br />
! scope="row" style="background: #7ac5e8" |Initial OD(t=0) <br />
! scope="row" style="background: #7ac5e8"|OD1 (t=1) <br />
! scope="row" style="background: #7ac5e8"|OD2 (t=1) <br />
! scope="row" style="background: #7ac5e8"|dilution used for measure t=1 <br />
! scope="row" style="background: #7ac5e8"|Mean OD <br />
! scope="row" style="background: #7ac5e8"|Growth Factor<br />
|-<br />
|01:01 <br />
|0,25 <br />
|0,32 <br />
|0,314 <br />
|10 <br />
|3,17 <br />
|12,68<br />
|-<br />
|09:01 <br />
|0,25 <br />
|0,148 <br />
|0,144 <br />
|10 <br />
|1,46 <br />
|5,84<br />
|-<br />
|01:09 <br />
|0,25 <br />
|0,138 <br />
|0,189 <br />
|10 <br />
|1,635 <br />
|6,54<br />
|-<br />
|01:01 <br />
|0,1 <br />
|0,109 <br />
|0,169 <br />
|10 <br />
|1,39 <br />
|13,9<br />
|-<br />
|09:01 <br />
|0,1 <br />
|0,04 <br />
|0,045 <br />
|10 <br />
|0,425 <br />
|4,25<br />
|-<br />
|01:09 <br />
|0,1 <br />
|0,067 <br />
|0,053 <br />
|10 <br />
|0,6 <br />
|6<br />
|-<br />
|01:01 <br />
|0,01 <br />
|0,067 <br />
|0,061 <br />
|1 <br />
|0,064 <br />
|6,4<br />
|-<br />
|09:01 <br />
|0,01 <br />
|0,056 <br />
|0,05 <br />
|1 <br />
|0,053 <br />
|5,3<br />
|-<br />
|01:09 <br />
|0,01 <br />
|0,074 <br />
|0,073 <br />
|1 <br />
|0,0735 <br />
|7,35<br />
|-<br />
|} <br />
<br />
<br />
{|<br />
|-<br />
|<!--column1-->[[File:HIS-BSAS2012.png|400px]]<br />
|}<br />
<br />
<br />
As shown in graph and table there is a basal growth that does not depend on the initial OD or strain proportion, of a growth factor of 6 approximately.<br />
However we observed a much higher growth at the proportion 1:1 when the initial OD 0.25 and 0.1. Therefore we can assume that at these proportions there is a natural cooperation between the strains and that should be the level of growth that we would like to assess through our bioengineering. Besides we would like to be able in the future to tune the strains in order to be able to obtain in the proportions 9:1 and 1:9 similar results to those obtained in the 1:1, at our own will.<br />
<br />
==== At the same initial OD: 0.2, followed over time ====<br />
<br />
We set the same cultures and cocultures as in point i), but starting all of them at the same OD: 0.2 and we followed them over 2 days. At day 1 we took pictures of them and at day 2 we measured the final OD. <br />
<br />
{| align="center" <br />
|- valign="top"<br />
|<br />
{| class="wikitable"<br />
|+ Cultures set at initial OD: 0.2 and measured over time (Days 0 and 2)<br />
! scope="row" style="background: #7ac5e8"|Strain<br />
! scope="row" style="background: #7ac5e8"|Day 0 <br />
! scope="row" style="background: #7ac5e8"|Day 2<br />
|-<br />
|TCY 3190 (-H+T) <br />
|0,2<br />
|2,92<br />
|-<br />
|TCY 3265 (+H-T) <br />
|0,2<br />
|0,19<br />
|-<br />
|Coculture of strains (TCY 3190- TCY 3265) <br />
|0,2<br />
|2,76<br />
|-<br />
|Negative control (TCY 3043 / -H-T) <br />
|0,2<br />
|0,6<br />
|-<br />
|Positive Control (TCY 3154/ +H+T) <br />
|0,2<br />
|2,54<br />
|}<br />
<br />
<br />
{| class="wikitable" border="1"<br />
|-<br />
|<!--column1-->[[File:Bsas2012-strains-wednesday.png|500px]]<br />
|-<br />
|Picture: Day 1 after starting cultures, shows different OD reached by strains. <br />
|}<br />
<br />
We repeated this experiment 4 times with different modifications: increasing the amount of days for up to a week, measuring every 12 hs instead of every 24 hs and using different strains. However, bacterial contaminations and the high rate of revertants prevented us from getting to a valid results in those cases, whereas the experiment up to day 2 always worked correctly. This denotes that we should assess the problem of contamination (for example including ampicilin in the cultures) and revertant rate (revising the design of the experiment or looking for more stable strains) as the impossibility to go further than day 2 may put limitations to some applications of the Synthetic Community.<br />
|}<br />
<br />
== Coculture in Agar and Revertant mutation control ==<br />
<br />
<br />
Through this experiment we aim to quantify the rate of revertants of each strain, and to asses if cross-feeding between a lawn of cells of one strain and colonies from and other strain is posible. <br />
<br />
We used petri dishes with agar medium with (+) and without (-) Trp and His as shown in the following table.<br />
<br />
We started a culture of each strain in synthetic complete medium, measured its OD 24 hs after the culture initiated, replaced the synthetic complete medium for one lacking both H and T (to avoid residual growth) and plated ~10^6 cells (lawn) or ~10^2 cells (seed) as shown by the following table (we considered OD600=1 represents 3*10^7 cells). <br />
At the same time, 3 controls (one for each strain) were carried in YPD complete medium to check the viability of each strain separately, and to estimate the seed CFU (colony formin units) more precisely. <br />
<br />
{| class="wikitable"<br />
! scope="row" style="background: #7ac5e8"|Medium H<br />
! scope="row" style="background: #7ac5e8"|Medium T<br />
! scope="row" style="background: #7ac5e8"|Lawn (10^6 cells) <br />
! scope="row" style="background: #7ac5e8"|Seed (10^2 cells) <br />
! scope="row" style="background: #7ac5e8"|Description of experiment <br />
! scope="row" style="background: #7ac5e8"|Results after 3 days - Replica 1<br />
! scope="row" style="background: #7ac5e8"|Results after 3 days - Replica 2<br />
|-<br />
|(-) <br />
|(+) <br />
|(-) <br />
|Strain –H+T <br />
|Control of His revertants <br />
|7 <br />
|7<br />
|-<br />
|-<br />
|(+)<br />
|(-)<br />
|(-)<br />
|Strain +H-T <br />
|Control of Trp revertants <br />
|2 <br />
|7<br />
|-<br />
|(-)<br />
|(-)<br />
|Strain +H-T<br />
|Strain –H+T<br />
|Coculture; we expect to see natural cooperation<br />
|960<br />
|800<br />
|-<br />
|(-)<br />
|(-)<br />
|Strain –H+T<br />
|Strain +H-T<br />
|Coculture; we expect to see natural cooperation<br />
|500<br />
|712<br />
|-<br />
|(-)<br />
|(-)<br />
|(-)<br />
|Strain +H+T<br />
|Viability of yeasts in medium<br />
|171<br />
|(-)<br />
|}<br />
<br />
'''Table: Shows description of each plate content and results in number of colonies counted by plate at day 3. YPD control results plates are not shown in the table'''. <br />
<br />
{| class="wikitable" border="1"<br />
|-<br />
|<!--column1-->[[File:Bsas2012-strains-placas2.jpg|300px]]<br />
|<!--column1-->[[File:Bsas2012-strains-placas1.jpg|300px]]<br />
|-<br />
|Petri Dishes<br />
| With marks of the counting of colonies<br />
|}<br />
<br />
<br />
==== Results ====<br />
The viability of the strains was high as expected, as well as the viability of a control positive strain in the –H-T medium. <br />
As shown in the table, we have a low, but existent, number of revertants from both his and trp auxotrophy strains. This number should be taken into account when interpreting the results from coculture growth after several days, given that the rate of revertants in liquid medium is probably the same. <br />
<br />
Growth in coculture was puzzling, as it resulted in more colonies than the expected. If cooperation was effective, we expected to see as many colonies as "seed" cells, not more. Revertion of cells from the "lawn" doesn't explain the number of colonies either. Probably a combination of both these effects are taking place.<br />
<br />
== Measurement of Trp in medium and Basal Production ==<br />
<br />
To check the efectiveness of our biobricks, we must first determine the ammount of tryptophan secreted by natural strains to the medium, so we can compare. With that end in mind, we designed a protocol for measurement of tryptophan in medium, based in its fluorescense at 350nm, when excited with 295nm light.<br />
As a previous step, we checked that none of the other aminoacids used in the medium interferes, by graphically comparing the spectres for uncomplemented medium and medium complemented with leucine, uracile and histidine, at an appropiate range.<br />
<br />
To determine Trp concentration, we must first have a way to transform our readings (intensity) to a more useful output, so we made a calibration curve, through serialized 1:2 dilutions of our medium, which Trp's concentration is 20μg/ml, until approximately constant intensity.<br />
<br />
The procedure to measure secretion rates will be growing the strain from a known OD in exponential growth phase in -T medium and plotting it's OD over time, spin-drying at time=t, retrieving the supernatant's Trp concentration and dividing it by the integral of OD vs. time between time=0 and time=t, so we get to a rate which will be proportional to the number of cells in the culture, which means we can actually compare between different strains. Since our medium is free from Trp, all of it should come from within the cells, and if the culture is growing at exponential rates, lysis should be negligible, so the only explanation would be cells exporting their own Trp.<br />
<br />
<br />
{| class="wikitable" border="1"<br />
|-<br />
|<!--column1-->[[File:Curva.png | 250px]]<br />
|-<br />
|Graph:Tryptophan calibration curve<br />
|}<br />
<br />
<br />
<br />
==== Results ====<br />
<br />
Through this experiment we can be sure that we would be able to measure increase of Trp in medium as it is exported from the cells, within the biological range of export.<br />
The sensitivity of this method seems to be enough to detect concentrations as low as ~0.01μg/ml, and as high as 20μg/ml, maybe more. Since our medium is 20μg/ml, we assume that's the saturation point of the curve. If we get bigger intensities than the one corresponding to it, we will dilute the sample.<br />
<br />
Because of time constraints, we haven't been able to check the method with either our designed strains nor the non-exporting ones.<br />
<br />
== Growth dependence on the Trp and His concentrations ==<br />
<br />
A important thing to characterize of the system is the dependence of the growth rate of the culture with the concentration of the crossfeeding aminoacids, tryptophane (Trp) and histidine (His). To do this we measured the final OD after an overnight growth in medium with different concentrations of Trp and His. <br />
<br />
We used strain ACL-379, that is auxotroph for both Trp and His. <br />
We prepared serial dilutions of SC medium in –T and –H medium, therefore creating two curves: one with decreasing concentrations of Trp and the other with decreasing concentrations of His. <br />
We then inoculated equal amounts of ACL-379 in each tube and incubated them overnight at 30°C with agitation. We took a picture of each tube and measured the OD600 reached by each culture.<br />
<br />
{| class="wikitable"<br />
|+Growth of ACL-379 as a function of Trp and His concentration<br />
! scope="row" style="background: #7ac5e8"|Medium<br />
! scope="row" style="background: #7ac5e8"|OD Replica 1<br />
! scope="row" style="background: #7ac5e8"|OD Replica 2<br />
|-<br />
|SC (no cells)<br />
|0,001<br />
|(-0,0036)<br />
|-<br />
| -T<br />
|(-0,003)<br />
|(-0,019)<br />
|-<br />
| Trp/2<br />
|2.56 <br />
|2.17<br />
|-<br />
| Trp/4<br />
|3.01 <br />
|3.11<br />
|-<br />
|Trp/8<br />
|1.54 <br />
|1.55<br />
|-<br />
|Trp/16<br />
|0.393 <br />
|0.409<br />
|-<br />
|Trp/32<br />
|0.013 <br />
|0.003<br />
|-<br />
| -H<br />
|(-0,008) <br />
|(-0,012)<br />
|-<br />
| His/2 <br />
|3.68 <br />
|3.84<br />
|-<br />
| His/4<br />
|2.07 <br />
|2.00<br />
|-<br />
|His/8<br />
|1.17 <br />
|0.97<br />
|-<br />
|His/16 <br />
|0.47 <br />
|0.432<br />
|-<br />
|His/32 <br />
|0.238 <br />
|0.257<br />
|-<br />
|SC (w/cells) <br />
|4.88 <br />
|4.91<br />
|} <br />
<br />
==== Results ====<br />
As expected the growth has a sigmoidal relationship with the concentration of Trp and His, when plotted in semilogarithmic scale. We call EC50 the effective concentration of each aminoacid at which the culture reaches 50% of the maximal growth. We considered these values as proxies of the Khis and Ktrp parameters of the [[Team:Buenos_Aires/Project/Model#Parameter_selection|mathematical model]], which can be used to estimate the secretion rate of each aminoacid needed to get effective crossfeeding. <br />
<br />
These results can also be observed by comparison of images that show the tubes at different OD. <br />
<br />
{| class="wikitable" border="1"<br />
|-<br />
|<!--column1-->[[File:Bsas2012-strains-alan1.png|250px]]<br />
|<!--column2-->[[File:Bsas2012-strains-ultima.jpg|250px]] <br />
|-<br />
|Images from HLU series<br />
|Images from TLU series<br />
|}<br />
<br />
Notes: <br />
SC: Synthetic complete medium with all the aminoacids. It was used as a blank for the spectrofluorometer.<br />
<br />
HTLU is the culture in the medium with all the required aminoacids.<br />
<br />
<br />
== Experimental determination of strains death rate==<br />
<br />
We set out to determine how long can auxotroph cells[link] survive in media that lacks both Trytophan and Histidine. These values are the '''death''' parameters for CFP and YFP strains used in our model[link]. These were taken as equal in the mathematical analysis for simplicity but now we would like to test whether this approximation is accurate.<br />
<br />
Given that our system most likely will present a lag phase until a certain amount of both AmioAcids is accumulated in the media, will the cells be viable until this occurs? This is a neccesary check of our '' system's feasability''.<br />
<br />
===== Protocol =====<br />
<br />
For this experiment we used<br />
{|<br />
|-<br />
|[[File:BsAs2012-icono-YFP.jpg|200px]]<br />
|[[File:BsAs2012-icono-CFP.jpg|200px]]<br />
|- align="center"<br />
|YFP Strain<br />
|CFP Strain<br />
|}<br />
<br />
*We set cultures of the two auxotroph strains without being transformed (YFP and CFP) in medium –HT at an initial OD of 0.01. <br />
*Each day we plated the same amount of µl of the culture and counted the number of colonies obtain in each plate. We set 3 replica of each strain.<br />
<br />
===== Result =====<br />
<br />
<br />
{| class="wikitable"<br />
! scope="row" style="background: #7ac5e8" |Strain<br />
! scope="row" style="background: #7ac5e8" |Replica<br />
! scope="row" style="background: #7ac5e8" |Monday<br />
! scope="row" style="background: #7ac5e8" |Tuesday<br />
! scope="row" style="background: #7ac5e8" |Wednesday<br />
|-<br />
|CFP<br />
|1<br />
|260<br />
|320<br />
|285<br />
|-<br />
|CFP<br />
|2<br />
|267<br />
|314<br />
|76<br />
|-<br />
|CFP<br />
|3<br />
|413<br />
|362<br />
|278<br />
|-<br />
|YFP<br />
|1<br />
|230<br />
|316<br />
|688<br />
|-<br />
|YFP<br />
|2<br />
|291<br />
|194<br />
|524<br />
|-<br />
|YFP<br />
|3<br />
|449<br />
|344<br />
|725<br />
|}<br />
<br />
'''Table:''' Number of colonies counted per plate.<br />
<br />
We expect to see a decrease in the number of colonies - because of cell death. We found that this was not the case, in the experiment's time lapse. However we observed that the size of the colonies was smaller everyday as can be seen in the following pictures.<br />
<br />
[[File:Bsas2012kdeathcells.png| 500px]]<br />
<br />
<br />
We can infer from this data that though they have not died, they may have enter into a '''...Alan state'''. In this way cells can survive for a period of time in media defficient in amino acid (at least, during the time course of our experiment), but grow slower. Probably this would require more time than 3 days to observe significative cell dying.<br />
<br />
<br />
{|<br />
|[[File:BsAs2012_celldeath.png | 100px]]<br />
|[[File:BsAs2012_celldeath2.png| 100px]]<br />
|<br />
|}<br />
FigureX.</div>Lucho.Morohttp://2012.igem.org/File:Curva.pngFile:Curva.png2012-10-27T03:26:16Z<p>Lucho.Moro: uploaded a new version of &quot;File:Curva.png&quot;</p>
<hr />
<div>Calibration curve</div>Lucho.Morohttp://2012.igem.org/File:Curva.pngFile:Curva.png2012-10-27T03:16:01Z<p>Lucho.Moro: Calibration curve</p>
<hr />
<div>Calibration curve</div>Lucho.Morohttp://2012.igem.org/Team:Buenos_Aires/HP/GarageLabTeam:Buenos Aires/HP/GarageLab2012-10-27T02:36:31Z<p>Lucho.Moro: /* Garage Lab */</p>
<hr />
<div>{{:Team:Buenos_Aires/Templates/menu}}<br />
<br />
= Garage Lab =<br />
{|<br />
|<br />
GarageLab is a group of scientists, artists and engineers cofounded by Emiliano Kargieman and Dario Wainer in 2009 as a Workshop about Bioengineering at the University of Palermo.<br />
<br />
During these years, it grew up to be a multitasking group, by getting involved in subjects such as Neuro-robotics, Contamination assessment, Data Privacy and Management, Personalized Medicine, Public Information and Problematics of Open Innovation.<br />
<br />
Now a days, the group has 22 active members and 5 projects with Non governmental organizations.<br />
<br />
They are evolving in order to become an institution and in process of building a proper infrastructure that would allow the resolution of high impact issues, as much as bigger workshops, hackatons, talks, conferences and developments. <br />
<br />
You can find out more about this creative and talented group of people at: http://garagelab.tumblr.com/ or their twitter http://twitter.com/garagelab<br />
|<br />
[[File:Bsas2012-Gl.png | 285px]]<br />
|}<br />
<br />
== Learn and play session ==<br />
{|<br />
|<br />
Of the several reunions that we had at Garage Lab, perhaps the most interesting was the one in which one of our members taught Synthetic Biology to a group of computer scientists and engineers through a game. The key question was not "How does DNA works?" but "How can we make it work for us?"<br />
<br />
Through these talks we find out how people from different disciplines other than natural sciences come up with new ideas and points of view that challenge and make us rethink biology.<br />
<br />
We are planning to develop a game (either a computer game, a cards game or both) for spreading synthetic biology concepts and strategies among young and non specialized people<br />
<br />
| width="30%" align="center" |<br />
{| style="width: 50%"<br />
| align="center" | [http://vimeo.com/45026664 [[File:Bsas2012-Garageee.jpg|220px]]]<br />
|-<br />
| style="text-align: center;" |[http://vimeo.com/45026664 Video]: SynBiology from a simple Computer Scientist point of view.<br />
|}<br />
<br />
|}<br />
<br />
== Future plans: Solving local problems ==<br />
{|<br />
| width="26%" | [[File:Bsas2012-Riachue.jpg|240px]]<br />
| At the beginning of 2012, Garage Lab contacted our instructors to begin a productive relationship. They believe in the impact the use of Synthetic Biology's standard parts could have in Argentina, as a tool to help solve endemic issues and they want it to become part of their projects and society. Since then we had a formal presentation of the group and our main project at a Garage Lab Meeting and several of our team members have attended the group meetings often. <br />
<br />
Shortly after the first contact with Garage Lab, they came to our laboratory and together we planned on giving use to the parts that come in the Kit in order to assess a key problem in Buenos Aires, which is the contamination present at Riachuelo, a branch of Río de la Plata. <br />
<br />
Several thousands of people live on the banks and nearby this highly polluted area which has been proven very hard to remediate from the environmental point of view, but also, a political challenge given its social and economic implications. <br />
|}<br />
<br />
{|<br />
|As a iGEM team we are thrilled to be part of this competition, but our ultimate goal is to spread the word about this discipline that is still ''new'' and ''unexplored'' here, not only to the general public but in the scientific community. Our collaboration with GarageLab will be a sort of testrun; a direct application of the ideas we've communicated about the Synthetic Biology Community and specifically the possibilities of iGEM developed free technology to help solve both big and small problems in our country. <br />
<br />
Therefore, we decided to explore the question Garage Lab presented us as a secondary project for after the Jamborees, to develop a way to identify and measure water contamination; such as heavy metals and maybe other noxious components at Riachuelo in the form of an unexpensive kit that could be distributed so that each community is able to periodically measure contamination levels in their area. '''The infrastructure is not present... ''''<br />
<br />
We aim to build a set of biosensors using parts from the Registry, more particularly the arsenic biosensor. We have already contacted Edinburgh 2006, Cambridge 2009 and SouthBend 2011 Teams regarding the procedures and experiences they have had related to arsenic detection using biobricks. We found an amazing support, ideas and encouragement from every team we contacted, including teachers and students.<br />
<br />
<br />
'''So, work will continue after the Jamborees and this is just the start!'''<br />
<br />
| [[File:Bsas2012-manu.jpg|240px]]<br />
|}<br />
<br />
= Impact in the media =<br />
<br />
"[http://www.eldia.com.ar/edis/20121022/alumnos-argentinos-primera-vez-mundial-biologia-informaciongeneral1.htm Alumnos argentinos, por primera vez en un “Mundial” de biología]" Diario el Día, 10/22/12.<br />
<br />
"[http://www.diariohoy.net/accion-verNota-id-213948-titulo-Argentinos_clasificaron_para_el_concurso_mundial_de_biolog%C3%ADa_sint%C3%A9tica_ Argentinos clasificaron para el concurso mundial de biología sintética]" Diario Hoy, 10/22/12.<br />
<br />
"[http://www.aptus.com.ar/noticia/2932-competencia-mundial-de-biologia-sintetica Competencia mundial de Biología Sintética]" Aptus Noticias educativas, 10/21/12.<br />
<br />
"[http://www.telam.com.ar/nota/41479/ Argentinos clasificaron para el concurso mundial de biología sintética]" Agencia de noticias TELAM, 10/20/12.<br />
<br />
"[http://www.tvperu.gob.pe/noticias/tecnologia/otros/38160-argentinos-clasificaron-para-el-concurso-mundial-de-biologia-sintetica.html Argentinos clasificaron para el concurso mundial de biología sintética]" TV Perú Noticias , 10/20/12.<br />
<br />
"[http://www.agenciacyta.org.ar/2012/10/debut-y-exito-argentino-en-olimpiadas-de-biologia-sintetica/ Debut y éxito argentino en olimpiadas de biología sintética]" Nota en el portal CyTA. 10/19/12.<br />
<br />
"[http://www.tomamateyavivate.com.ar/formacion-y-universidades-argentinas/debut-y-exito-argentino-en-olimpiadas-de-biologia-sintetica/ Debut y éxito argentino en olimpiadas de biología sintética]" Tomamateyavivate, 10/19/12.<br />
<br />
"[http://www.conicet.gov.ar/?p=2827 Estudiantes argentinos pasan a la etapa final de un certamen mundial]" Nota en la página de noticias del CONICET, 10/19/2012.<br />
<br />
"[http://www.pagina12.com.ar/diario/sociedad/3-206038-2012-10-21.html Elegidos del Conicet]" Página 12, 20/10/12.<br />
<br />
"[http://blogs.scientificamerican.com/lab-rat/2012/09/09/igem-buenos-aires-synthetic-bacterial-communities/ iGEM Buenos Aires: Synthetic bacterial communities]" Lab Rat blog at scientificamerican.com 9/9/12.</div>Lucho.Morohttp://2012.igem.org/Team:Buenos_Aires/HP/GarageLabTeam:Buenos Aires/HP/GarageLab2012-10-27T02:30:35Z<p>Lucho.Moro: /* Future plans: Solving local problems */</p>
<hr />
<div>{{:Team:Buenos_Aires/Templates/menu}}<br />
<br />
= Garage Lab =<br />
{|<br />
|<br />
GarageLab is a group of scientists, artists and engineers cofounded by Emiliano Kargieman and Dario Wainer in 2009 as a Workshop about Bioengineering at the University of Palermo.<br />
<br />
During these years, it grew up to be a multitasking group, by getting involved in subjects such as Neuro-robotics, Contamination assessment, Data Privacy and Management, Personalized Medicine, Public Information and Problematics of Open Innovation.<br />
<br />
Now a days, the group has 22 active members and 5 projects with Non governmental organizations.<br />
<br />
They are evolving in order to become an institution and in process of building a propper infrastructure that would allow the resolution of high impact issues, as much as bigger workshops, hackatons, talks, conferences and developments. <br />
<br />
You can find out more about this creative and talented group of people at: http://garagelab.tumblr.com/ or their twitter http://twitter.com/garagelab<br />
|<br />
[[File:Bsas2012-Gl.png | 285px]]<br />
|}<br />
<br />
== Learn and play session ==<br />
{|<br />
|<br />
Of the several reunions that we had at Garage Lab, perhaps the most interesting was the one in which one of our members taught Synthetic Biology to a group of computer scientists and engineers through a game. The key question was not "How does DNA works?" but "How can we make it work for us?"<br />
<br />
Through these talks we find out how people from different disciplines other than natural sciences come up with new ideas and points of view that challenge and make us rethink biology.<br />
<br />
We are planing to develop a game (either a computer game, a cards game or both) for spreading synthetic biology concepts and strategies among young and non specialized people<br />
<br />
| width="30%" align="center" |<br />
{| style="width: 50%"<br />
| align="center" | [http://vimeo.com/45026664 [[File:Bsas2012-Garageee.jpg|220px]]]<br />
|-<br />
| style="text-align: center;" |[http://vimeo.com/45026664 Video]: SynBiology from a simple Computer Scientist point of view.<br />
|}<br />
<br />
|}<br />
<br />
== Future plans: Solving local problems ==<br />
{|<br />
| width="26%" | [[File:Bsas2012-Riachue.jpg|240px]]<br />
| At the beginning of 2012, Garage Lab contacted our instructors to begin a productive relationship. They believe in the impact the use of Synthetic Biology's standard parts could have in Argentina, as a tool to help solve endemic issues and they want it to become part of their projects and society. Since then we had a formal presentation of the group and our main project at a Garage Lab Meeting and several of our team members have attended the group meetings often. <br />
<br />
Shortly after the first contact with Garage Lab, they came to our laboratory and together we planned on giving use to the parts that come in the Kit in order to assess a key problem in Buenos Aires, which is the contamination present at Riachuelo, a branch of Río de la Plata. <br />
<br />
Several thousands of people live on the banks and nearby this highly polluted area which has been proven very hard to remediate from the environmental point of view, but also, a political challenge given its social and economic implications. <br />
|}<br />
<br />
{|<br />
|As a iGEM team we are thrilled to be part of this competition, but our ultimate goal is to spread the word about this discipline that is still ''new'' and ''unexplored'' here, not only to the general public but in the scientific community. Our collaboration with GarageLab will be a sort of testrun; a direct application of the ideas we've communicated about the Synthetic Biology Community and specifically the possibilities of iGEM developed free technology to help solve both big and small problems in our country. <br />
<br />
Therefore, we decided to explore the question Garage Lab presented us as a secondary project for after the Jamborees, to develop a way to identify and measure water contamination; such as heavy metals and maybe other noxious components at Riachuelo in the form of an unexpensive kit that could be distributed so that each community is able to periodically measure contamination levels in their area. '''The infrastructure is not present... ''''<br />
<br />
We aim to build a set of biosensors using parts from the Registry, more particulary the arsenic biosensor. We have already contacted Edinburgh 2006, Cambridge 2009 and SouthBend 2011 Teams regarding the procedures and experiences they have had related to arsenic detection using biobricks. We found an amazing support, ideas and encouragement from every team we contacted, including teachers and students.<br />
<br />
<br />
'''So, work will continue after the Jamborees and this is just the start!'''<br />
<br />
| [[File:Bsas2012-manu.jpg|240px]]<br />
|}<br />
<br />
= Impact in the media =<br />
<br />
"[http://www.eldia.com.ar/edis/20121022/alumnos-argentinos-primera-vez-mundial-biologia-informaciongeneral1.htm Alumnos argentinos, por primera vez en un “Mundial” de biología]" Diario el Día, 10/22/12.<br />
<br />
"[http://www.diariohoy.net/accion-verNota-id-213948-titulo-Argentinos_clasificaron_para_el_concurso_mundial_de_biolog%C3%ADa_sint%C3%A9tica_ Argentinos clasificaron para el concurso mundial de biología sintética]" Diario Hoy, 10/22/12.<br />
<br />
"[http://www.aptus.com.ar/noticia/2932-competencia-mundial-de-biologia-sintetica Competencia mundial de Biología Sintética]" Aptus Noticias educativas, 10/21/12.<br />
<br />
"[http://www.telam.com.ar/nota/41479/ Argentinos clasificaron para el concurso mundial de biología sintética]" Agencia de noticias TELAM, 10/20/12.<br />
<br />
"[http://www.tvperu.gob.pe/noticias/tecnologia/otros/38160-argentinos-clasificaron-para-el-concurso-mundial-de-biologia-sintetica.html Argentinos clasificaron para el concurso mundial de biología sintética]" TV Perú Noticias , 10/20/12.<br />
<br />
"[http://www.agenciacyta.org.ar/2012/10/debut-y-exito-argentino-en-olimpiadas-de-biologia-sintetica/ Debut y éxito argentino en olimpiadas de biología sintética]" Nota en el portal CyTA. 10/19/12.<br />
<br />
"[http://www.tomamateyavivate.com.ar/formacion-y-universidades-argentinas/debut-y-exito-argentino-en-olimpiadas-de-biologia-sintetica/ Debut y éxito argentino en olimpiadas de biología sintética]" Tomamateyavivate, 10/19/12.<br />
<br />
"[http://www.conicet.gov.ar/?p=2827 Estudiantes argentinos pasan a la etapa final de un certamen mundial]" Nota en la página de noticias del CONICET, 10/19/2012.<br />
<br />
"[http://www.pagina12.com.ar/diario/sociedad/3-206038-2012-10-21.html Elegidos del Conicet]" Página 12, 20/10/12.<br />
<br />
"[http://blogs.scientificamerican.com/lab-rat/2012/09/09/igem-buenos-aires-synthetic-bacterial-communities/ iGEM Buenos Aires: Synthetic bacterial communities]" Lab Rat blog at scientificamerican.com 9/9/12.</div>Lucho.Morohttp://2012.igem.org/Team:Buenos_Aires/HP/GarageLabTeam:Buenos Aires/HP/GarageLab2012-10-27T02:26:18Z<p>Lucho.Moro: /* Future plans: Solving local problems */</p>
<hr />
<div>{{:Team:Buenos_Aires/Templates/menu}}<br />
<br />
= Garage Lab =<br />
{|<br />
|<br />
GarageLab is a group of scientists, artists and engineers cofounded by Emiliano Kargieman and Dario Wainer in 2009 as a Workshop about Bioengineering at the University of Palermo.<br />
<br />
During these years, it grew up to be a multitasking group, by getting involved in subjects such as Neuro-robotics, Contamination assessment, Data Privacy and Management, Personalized Medicine, Public Information and Problematics of Open Innovation.<br />
<br />
Now a days, the group has 22 active members and 5 projects with Non governmental organizations.<br />
<br />
They are evolving in order to become an institution and in process of building a propper infrastructure that would allow the resolution of high impact issues, as much as bigger workshops, hackatons, talks, conferences and developments. <br />
<br />
You can find out more about this creative and talented group of people at: http://garagelab.tumblr.com/ or their twitter http://twitter.com/garagelab<br />
|<br />
[[File:Bsas2012-Gl.png | 285px]]<br />
|}<br />
<br />
== Learn and play session ==<br />
{|<br />
|<br />
Of the several reunions that we had at Garage Lab, perhaps the most interesting was the one in which one of our members taught Synthetic Biology to a group of computer scientists and engineers through a game. The key question was not "How does DNA works?" but "How can we make it work for us?"<br />
<br />
Through these talks we find out how people from different disciplines other than natural sciences come up with new ideas and points of view that challenge and make us rethink biology.<br />
<br />
We are planing to develop a game (either a computer game, a cards game or both) for spreading synthetic biology concepts and strategies among young and non specialized people<br />
<br />
| width="30%" align="center" |<br />
{| style="width: 50%"<br />
| align="center" | [http://vimeo.com/45026664 [[File:Bsas2012-Garageee.jpg|220px]]]<br />
|-<br />
| style="text-align: center;" |[http://vimeo.com/45026664 Video]: SynBiology from a simple Computer Scientist point of view.<br />
|}<br />
<br />
|}<br />
<br />
== Future plans: Solving local problems ==<br />
{|<br />
| width="26%" | [[File:Bsas2012-Riachue.jpg|240px]]<br />
| At the beginning of 2012, Garage Lab contacted our instructors to begin a productive relationship. They believe in the impact the use of Synthetic Biology's standard parts of could have in Argentina, as a tool to help solve endemic issues and they want it to become part of their projects and society. Since then we had a formal presentation of the group and our main project at a Garage Lab Meeting and several of our team members have attended the group meetings often. <br />
<br />
Shortly after the first contact with Garage Lab, they came to our laboratory and together we planned on giving use to the parts that come in the Kit in order to assess a key problem in Buenos Aires, which is the contamination present at Riachuelo, a branch of Río de la Plata. <br />
<br />
Several thousands of people live on the banks and nearby this highly polluted area which has been proven very hard to remediate from the environmental point of view, but also, a political challenge given its social and economic implications. <br />
|}<br />
<br />
{|<br />
|As a iGEM team we are thrilled to be part of this competition, but our ultimate goal is to spread the word about this discipline that it's still ''new'' and ''unexplored'' here, not only to the general public but in the scientific comunnity. Our collaboration with GarageLab will be a sort of testrun;a direct application of the ideas we've comunicated about the Synthetic Biology Community and specifically the possibilities of iGEM developed free technology to help solve both big and small problems in our country. <br />
<br />
Therefore, we decided to explore the question Garage Lab presented us as a secondary project for after the Jamborees, to develop a way to identify and measure water contamination; such as heavy metals and maybe other noxious components at Riachuelo in the form of an unexpensive kit that could be distributed so that each community is able to periodically measure contamination levels in their area. '''The infrastructure is not present... ''''<br />
<br />
We aim to build a set of biosensors using the parts from the Registry, more particulary the arsenic biosensor. We have already contacted Edinburgh 2006, Cambridge 2009 and SouthBend 2011 Teams regarding the procedures and experiences the have had related to arsenic detection using biobricks. We found an amazing support, ideas and encouragement from every team we contacted, including teachers and students.<br />
<br />
<br />
'''So, work will continue after the Jamborees and this is just the start!'''<br />
<br />
| [[File:Bsas2012-manu.jpg|240px]]<br />
|}<br />
<br />
= Impact in the media =<br />
<br />
"[http://www.eldia.com.ar/edis/20121022/alumnos-argentinos-primera-vez-mundial-biologia-informaciongeneral1.htm Alumnos argentinos, por primera vez en un “Mundial” de biología]" Diario el Día, 10/22/12.<br />
<br />
"[http://www.diariohoy.net/accion-verNota-id-213948-titulo-Argentinos_clasificaron_para_el_concurso_mundial_de_biolog%C3%ADa_sint%C3%A9tica_ Argentinos clasificaron para el concurso mundial de biología sintética]" Diario Hoy, 10/22/12.<br />
<br />
"[http://www.aptus.com.ar/noticia/2932-competencia-mundial-de-biologia-sintetica Competencia mundial de Biología Sintética]" Aptus Noticias educativas, 10/21/12.<br />
<br />
"[http://www.telam.com.ar/nota/41479/ Argentinos clasificaron para el concurso mundial de biología sintética]" Agencia de noticias TELAM, 10/20/12.<br />
<br />
"[http://www.tvperu.gob.pe/noticias/tecnologia/otros/38160-argentinos-clasificaron-para-el-concurso-mundial-de-biologia-sintetica.html Argentinos clasificaron para el concurso mundial de biología sintética]" TV Perú Noticias , 10/20/12.<br />
<br />
"[http://www.agenciacyta.org.ar/2012/10/debut-y-exito-argentino-en-olimpiadas-de-biologia-sintetica/ Debut y éxito argentino en olimpiadas de biología sintética]" Nota en el portal CyTA. 10/19/12.<br />
<br />
"[http://www.tomamateyavivate.com.ar/formacion-y-universidades-argentinas/debut-y-exito-argentino-en-olimpiadas-de-biologia-sintetica/ Debut y éxito argentino en olimpiadas de biología sintética]" Tomamateyavivate, 10/19/12.<br />
<br />
"[http://www.conicet.gov.ar/?p=2827 Estudiantes argentinos pasan a la etapa final de un certamen mundial]" Nota en la página de noticias del CONICET, 10/19/2012.<br />
<br />
"[http://www.pagina12.com.ar/diario/sociedad/3-206038-2012-10-21.html Elegidos del Conicet]" Página 12, 20/10/12.<br />
<br />
"[http://blogs.scientificamerican.com/lab-rat/2012/09/09/igem-buenos-aires-synthetic-bacterial-communities/ iGEM Buenos Aires: Synthetic bacterial communities]" Lab Rat blog at scientificamerican.com 9/9/12.</div>Lucho.Morohttp://2012.igem.org/Team:Buenos_Aires/HP/GarageLabTeam:Buenos Aires/HP/GarageLab2012-10-27T02:25:20Z<p>Lucho.Moro: /* Learn and play session */</p>
<hr />
<div>{{:Team:Buenos_Aires/Templates/menu}}<br />
<br />
= Garage Lab =<br />
{|<br />
|<br />
GarageLab is a group of scientists, artists and engineers cofounded by Emiliano Kargieman and Dario Wainer in 2009 as a Workshop about Bioengineering at the University of Palermo.<br />
<br />
During these years, it grew up to be a multitasking group, by getting involved in subjects such as Neuro-robotics, Contamination assessment, Data Privacy and Management, Personalized Medicine, Public Information and Problematics of Open Innovation.<br />
<br />
Now a days, the group has 22 active members and 5 projects with Non governmental organizations.<br />
<br />
They are evolving in order to become an institution and in process of building a propper infrastructure that would allow the resolution of high impact issues, as much as bigger workshops, hackatons, talks, conferences and developments. <br />
<br />
You can find out more about this creative and talented group of people at: http://garagelab.tumblr.com/ or their twitter http://twitter.com/garagelab<br />
|<br />
[[File:Bsas2012-Gl.png | 285px]]<br />
|}<br />
<br />
== Learn and play session ==<br />
{|<br />
|<br />
Of the several reunions that we had at Garage Lab, perhaps the most interesting was the one in which one of our members taught Synthetic Biology to a group of computer scientists and engineers through a game. The key question was not "How does DNA works?" but "How can we make it work for us?"<br />
<br />
Through these talks we find out how people from different disciplines other than natural sciences come up with new ideas and points of view that challenge and make us rethink biology.<br />
<br />
We are planing to develop a game (either a computer game, a cards game or both) for spreading synthetic biology concepts and strategies among young and non specialized people<br />
<br />
| width="30%" align="center" |<br />
{| style="width: 50%"<br />
| align="center" | [http://vimeo.com/45026664 [[File:Bsas2012-Garageee.jpg|220px]]]<br />
|-<br />
| style="text-align: center;" |[http://vimeo.com/45026664 Video]: SynBiology from a simple Computer Scientist point of view.<br />
|}<br />
<br />
|}<br />
<br />
== Future plans: Solving local problems ==<br />
{|<br />
| width="26%" | [[File:Bsas2012-Riachue.jpg|240px]]<br />
| At the beggining of 2012, Garage Lab contacted our instructors to begin a productive relationship. They believe in the impact the use of Synthetic Biology's standard parts of could have in Argentina, as a tool to help solve endemic issues and they want it to become part of their projects and society. Since then we had a formal presentation of the group and our main project at a Garage Lab Meeting and several of our team members have attended the group meetings often. <br />
<br />
Shortly after the first contact with Garage Lab, they came to our laboratory and together we planned on giving use to the parts that come in the Kit in order to assess a key problem in Buenos Aires, which is the contamination present at Riachuelo, a branch of Río de la Plata. <br />
<br />
Several thousands of people live on the banks and nearby this highly polluted area which has been proven very hard to remediate from the environmental point of view, but also, a political challenge given its social and economic implications. <br />
|}<br />
<br />
{|<br />
|As a iGEM team we are thrilled to be part of this competition, but our ultimate goal is to spread the word about this discipline that it's still ''new'' and ''unexplored'' here, not only to the general public but in the scientific comunnity. Our collaboration with GarageLab will be a sort of testrun;a direct application of the ideas we've comunicated about the Synthetic Biology Community and specifically the possibilities of iGEM developed free technology to help solve both big and small problems in our country. <br />
<br />
Therefore, we decided to explore the question Garage Lab presented us as a secondary project for after the Jamborees, to develop a way to identify and measure water contamination; such as heavy metals and maybe other noxious components at Riachuelo in the form of an unexpensive kit that could be distributed so that each community is able to periodically measure contamination levels in their area. '''The infrastructure is not present... ''''<br />
<br />
We aim to build a set of biosensors using the parts from the Registry, more particulary the arsenic biosensor. We have already contacted Edinburgh 2006, Cambridge 2009 and SouthBend 2011 Teams regarding the procedures and experiences the have had related to arsenic detection using biobricks. We found an amazing support, ideas and encouragement from every team we contacted, including teachers and students.<br />
<br />
<br />
'''So, work will continue after the Jamborees and this is just the start!'''<br />
<br />
| [[File:Bsas2012-manu.jpg|240px]]<br />
|}<br />
<br />
= Impact in the media =<br />
<br />
"[http://www.eldia.com.ar/edis/20121022/alumnos-argentinos-primera-vez-mundial-biologia-informaciongeneral1.htm Alumnos argentinos, por primera vez en un “Mundial” de biología]" Diario el Día, 10/22/12.<br />
<br />
"[http://www.diariohoy.net/accion-verNota-id-213948-titulo-Argentinos_clasificaron_para_el_concurso_mundial_de_biolog%C3%ADa_sint%C3%A9tica_ Argentinos clasificaron para el concurso mundial de biología sintética]" Diario Hoy, 10/22/12.<br />
<br />
"[http://www.aptus.com.ar/noticia/2932-competencia-mundial-de-biologia-sintetica Competencia mundial de Biología Sintética]" Aptus Noticias educativas, 10/21/12.<br />
<br />
"[http://www.telam.com.ar/nota/41479/ Argentinos clasificaron para el concurso mundial de biología sintética]" Agencia de noticias TELAM, 10/20/12.<br />
<br />
"[http://www.tvperu.gob.pe/noticias/tecnologia/otros/38160-argentinos-clasificaron-para-el-concurso-mundial-de-biologia-sintetica.html Argentinos clasificaron para el concurso mundial de biología sintética]" TV Perú Noticias , 10/20/12.<br />
<br />
"[http://www.agenciacyta.org.ar/2012/10/debut-y-exito-argentino-en-olimpiadas-de-biologia-sintetica/ Debut y éxito argentino en olimpiadas de biología sintética]" Nota en el portal CyTA. 10/19/12.<br />
<br />
"[http://www.tomamateyavivate.com.ar/formacion-y-universidades-argentinas/debut-y-exito-argentino-en-olimpiadas-de-biologia-sintetica/ Debut y éxito argentino en olimpiadas de biología sintética]" Tomamateyavivate, 10/19/12.<br />
<br />
"[http://www.conicet.gov.ar/?p=2827 Estudiantes argentinos pasan a la etapa final de un certamen mundial]" Nota en la página de noticias del CONICET, 10/19/2012.<br />
<br />
"[http://www.pagina12.com.ar/diario/sociedad/3-206038-2012-10-21.html Elegidos del Conicet]" Página 12, 20/10/12.<br />
<br />
"[http://blogs.scientificamerican.com/lab-rat/2012/09/09/igem-buenos-aires-synthetic-bacterial-communities/ iGEM Buenos Aires: Synthetic bacterial communities]" Lab Rat blog at scientificamerican.com 9/9/12.</div>Lucho.Morohttp://2012.igem.org/Team:Buenos_Aires/HP/GarageLabTeam:Buenos Aires/HP/GarageLab2012-10-27T02:21:12Z<p>Lucho.Moro: /* Impact in the media */</p>
<hr />
<div>{{:Team:Buenos_Aires/Templates/menu}}<br />
<br />
= Garage Lab =<br />
{|<br />
|<br />
GarageLab is a group of scientists, artists and engineers cofounded by Emiliano Kargieman and Dario Wainer in 2009 as a Workshop about Bioengineering at the University of Palermo.<br />
<br />
During these years, it grew up to be a multitasking group, by getting involved in subjects such as Neuro-robotics, Contamination assessment, Data Privacy and Management, Personalized Medicine, Public Information and Problematics of Open Innovation.<br />
<br />
Now a days, the group has 22 active members and 5 projects with Non governmental organizations.<br />
<br />
They are evolving in order to become an institution and in process of building a propper infrastructure that would allow the resolution of high impact issues, as much as bigger workshops, hackatons, talks, conferences and developments. <br />
<br />
You can find out more about this creative and talented group of people at: http://garagelab.tumblr.com/ or their twitter http://twitter.com/garagelab<br />
|<br />
[[File:Bsas2012-Gl.png | 285px]]<br />
|}<br />
<br />
== Learn and play session ==<br />
{|<br />
|<br />
Of the several reunions that we had at Garage Lab, perhaps the most interesting was the one in which one of our members taught Synthetic Biology to a group of computer scientists and engineers through a game. The key question was not how does DNA works but How can we make it work for us?<br />
<br />
Through these talks we find out how people from different disciplines other than natural sciences come up with new ideas and points of view that challenge and make us rethink biology.<br />
<br />
We are planing to develop a game (either a computer game, a cards game or both) for spreading synthetic biology concepts and strategies among young and non specialized people<br />
<br />
| width="30%" align="center" |<br />
{| style="width: 50%"<br />
| align="center" | [http://vimeo.com/45026664 [[File:Bsas2012-Garageee.jpg|220px]]]<br />
|-<br />
| style="text-align: center;" |[http://vimeo.com/45026664 Video]: SynBiology from a simple Computer Scientist point of view.<br />
|}<br />
<br />
|}<br />
<br />
<br />
== Future plans: Solving local problems ==<br />
{|<br />
| width="26%" | [[File:Bsas2012-Riachue.jpg|240px]]<br />
| At the beggining of 2012, Garage Lab contacted our instructors to begin a productive relationship. They believe in the impact the use of Synthetic Biology's standard parts of could have in Argentina, as a tool to help solve endemic issues and they want it to become part of their projects and society. Since then we had a formal presentation of the group and our main project at a Garage Lab Meeting and several of our team members have attended the group meetings often. <br />
<br />
Shortly after the first contact with Garage Lab, they came to our laboratory and together we planned on giving use to the parts that come in the Kit in order to assess a key problem in Buenos Aires, which is the contamination present at Riachuelo, a branch of Río de la Plata. <br />
<br />
Several thousands of people live on the banks and nearby this highly polluted area which has been proven very hard to remediate from the environmental point of view, but also, a political challenge given its social and economic implications. <br />
|}<br />
<br />
{|<br />
|As a iGEM team we are thrilled to be part of this competition, but our ultimate goal is to spread the word about this discipline that it's still ''new'' and ''unexplored'' here, not only to the general public but in the scientific comunnity. Our collaboration with GarageLab will be a sort of testrun;a direct application of the ideas we've comunicated about the Synthetic Biology Community and specifically the possibilities of iGEM developed free technology to help solve both big and small problems in our country. <br />
<br />
Therefore, we decided to explore the question Garage Lab presented us as a secondary project for after the Jamborees, to develop a way to identify and measure water contamination; such as heavy metals and maybe other noxious components at Riachuelo in the form of an unexpensive kit that could be distributed so that each community is able to periodically measure contamination levels in their area. '''The infrastructure is not present... ''''<br />
<br />
We aim to build a set of biosensors using the parts from the Registry, more particulary the arsenic biosensor. We have already contacted Edinburgh 2006, Cambridge 2009 and SouthBend 2011 Teams regarding the procedures and experiences the have had related to arsenic detection using biobricks. We found an amazing support, ideas and encouragement from every team we contacted, including teachers and students.<br />
<br />
<br />
'''So, work will continue after the Jamborees and this is just the start!'''<br />
<br />
| [[File:Bsas2012-manu.jpg|240px]]<br />
|}<br />
<br />
= Impact in the media =<br />
<br />
"[http://www.eldia.com.ar/edis/20121022/alumnos-argentinos-primera-vez-mundial-biologia-informaciongeneral1.htm Alumnos argentinos, por primera vez en un “Mundial” de biología]" Diario el Día, 10/22/12.<br />
<br />
"[http://www.diariohoy.net/accion-verNota-id-213948-titulo-Argentinos_clasificaron_para_el_concurso_mundial_de_biolog%C3%ADa_sint%C3%A9tica_ Argentinos clasificaron para el concurso mundial de biología sintética]" Diario Hoy, 10/22/12.<br />
<br />
"[http://www.aptus.com.ar/noticia/2932-competencia-mundial-de-biologia-sintetica Competencia mundial de Biología Sintética]" Aptus Noticias educativas, 10/21/12.<br />
<br />
"[http://www.telam.com.ar/nota/41479/ Argentinos clasificaron para el concurso mundial de biología sintética]" Agencia de noticias TELAM, 10/20/12.<br />
<br />
"[http://www.tvperu.gob.pe/noticias/tecnologia/otros/38160-argentinos-clasificaron-para-el-concurso-mundial-de-biologia-sintetica.html Argentinos clasificaron para el concurso mundial de biología sintética]" TV Perú Noticias , 10/20/12.<br />
<br />
"[http://www.agenciacyta.org.ar/2012/10/debut-y-exito-argentino-en-olimpiadas-de-biologia-sintetica/ Debut y éxito argentino en olimpiadas de biología sintética]" Nota en el portal CyTA. 10/19/12.<br />
<br />
"[http://www.tomamateyavivate.com.ar/formacion-y-universidades-argentinas/debut-y-exito-argentino-en-olimpiadas-de-biologia-sintetica/ Debut y éxito argentino en olimpiadas de biología sintética]" Tomamateyavivate, 10/19/12.<br />
<br />
"[http://www.conicet.gov.ar/?p=2827 Estudiantes argentinos pasan a la etapa final de un certamen mundial]" Nota en la página de noticias del CONICET, 10/19/2012.<br />
<br />
"[http://www.pagina12.com.ar/diario/sociedad/3-206038-2012-10-21.html Elegidos del Conicet]" Página 12, 20/10/12.<br />
<br />
"[http://blogs.scientificamerican.com/lab-rat/2012/09/09/igem-buenos-aires-synthetic-bacterial-communities/ iGEM Buenos Aires: Synthetic bacterial communities]" Lab Rat blog at scientificamerican.com 9/9/12.</div>Lucho.Morohttp://2012.igem.org/Team:Buenos_AiresTeam:Buenos Aires2012-10-27T02:20:26Z<p>Lucho.Moro: /* Welcome to Buenos Aires 2012 iGEM Wiki! */</p>
<hr />
<div>{{:Team:Buenos_Aires/Templates/menu}}<br />
<br />
= Welcome to Buenos Aires 2012 iGEM Wiki! =<br />
<br />
{|<br />
|<br />
'''What are you looking for?'''<br />
<br />
* Check the [[Team:Buenos_Aires/Safety|safety questions]] and [[ Team:Buenos_Aires/Attributions | attributions]].<br />
<br />
* Meet [[Team:Buenos_Aires/Team/Members | the team]]. You can also read a little about [[Team:Buenos_Aires/Team/BsAs | where we come from]].<br />
<br />
* Learn about [[Team:Buenos_Aires/Project | our project]] (and don't forget to check [[Team:Buenos_Aires/Project/Schemes | all the schemes]] we thought to solve the problem).<br />
<br />
* Don't miss our yeast [[Team:Buenos_Aires/Results/Strains| strains characterization]], and the [[Team:Buenos_Aires/Results/Bb1 |main biobricks and devices ]] we designed and added to the registry. You can also find information about our ''planB'' [[Team:Buenos_Aires/Results/Bb2 | backup biobrick]] ''(aka. thank-you-customs biobrick)'' .<br />
<br />
* Take a look at our [[Team:Buenos_Aires/Project/Model | mathematical model]] of a synthetic ecology. And if you dare, take a look at the [[Team:Buenos_Aires/Project/ModelAdvance | advanced model]].<br />
<br />
* What we did outside the lab ''(aka. human practices)'' to [[Team:Buenos_Aires/HP/GarageLab | teach what synBio is about]], [[Team:Buenos_Aires/HP/GarageLab | start solving local problems ]] and [[ Team:Buenos_Aires/HP/EMBO | seed SynBio in Latin America]].<br />
<br />
* Watch an [http://youtu.be/bkczB60RziU online presentation] of our team and our project.<br />
<br />
* If you understand spanish, you can also check our [http://www.youtube.com/watch?v=oEMXc6cmmgo presentation video].<br />
<br />
* See the [https://2012.igem.org/Team:Buenos_Aires/HP/GarageLab#Impact_in_the_media impact in local newspapers].<br />
<br />
|<br />
{| style="width: 100%"<br />
| align="center" | [[File:Bsas2012-The Team.png|400px]]<br />
|}<br />
<br />
|}</div>Lucho.Morohttp://2012.igem.org/Team:Buenos_AiresTeam:Buenos Aires2012-10-27T02:06:37Z<p>Lucho.Moro: /* Welcome to Buenos Aires 2012 iGEM Wiki! */</p>
<hr />
<div>{{:Team:Buenos_Aires/Templates/menu}}<br />
<br />
= Welcome to Buenos Aires 2012 iGEM Wiki! =<br />
<br />
{|<br />
|<br />
'''What are you looking for?'''<br />
<br />
* Check the [[Team:Buenos_Aires/Safety|safety questions]] and [[ Team:Buenos_Aires/Attributions | attributions]].<br />
<br />
* Meet [[Team:Buenos_Aires/Team/Members | the team]]. You can also read a little about [[Team:Buenos_Aires/Team/BsAs | where we come from]].<br />
<br />
* Learn about [[Team:Buenos_Aires/Project | our project]] (and don't forget to check [[Team:Buenos_Aires/Project/Schemes | all the schemes]] we thought to solve the problem).<br />
<br />
* Don't miss our yeast [[Team:Buenos_Aires/Results/Strains| strains characterization]], and the [[Team:Buenos_Aires/Results/Bb1 |main biobricks and devices ]] we designed and added to the registry. You can also find information about our ''planB'' [[Team:Buenos_Aires/Results/Bb2 | backup biobrick]] ''(aka. thank-you-customs biobrick)'' .<br />
<br />
* Take a look at our [[Team:Buenos_Aires/Project/Model | mathematical model]] of a synthetic ecology. And if you dare, take a look at the [[Team:Buenos_Aires/Project/ModelAdvance | advanced model]].<br />
<br />
* What we did outside the lab ''(aka. human practices)'' to [[Team:Buenos_Aires/HP/GarageLab | teach what synBio is about]], [[Team:Buenos_Aires/HP/GarageLab | start solving local problems ]] and [[ Team:Buenos_Aires/HP/EMBO | seed SynBio in Latin America]].<br />
<br />
* Watch an [http://youtu.be/bkczB60RziU online presentation] of our team and our project.<br />
<br />
* If you understand spanish, you can also check our [http://www.youtube.com/watch?v=oEMXc6cmmgo presentation video].<br />
<br />
<br />
|<br />
{| style="width: 100%"<br />
| align="center" | [[File:Bsas2012-The Team.png|400px]]<br />
|}<br />
<br />
|}</div>Lucho.Morohttp://2012.igem.org/Team:Buenos_Aires/HP/GarageLabTeam:Buenos Aires/HP/GarageLab2012-10-27T02:05:20Z<p>Lucho.Moro: /* Impact in the media */</p>
<hr />
<div>{{:Team:Buenos_Aires/Templates/menu}}<br />
<br />
= Garage Lab =<br />
{|<br />
|<br />
GarageLab is a group of scientists, artists and engineers cofounded by Emiliano Kargieman and Dario Wainer in 2009 as a Workshop about Bioengineering at the University of Palermo.<br />
<br />
During these years, it grew up to be a multitasking group, by getting involved in subjects such as Neuro-robotics, Contamination assessment, Data Privacy and Management, Personalized Medicine, Public Information and Problematics of Open Innovation.<br />
<br />
Now a days, the group has 22 active members and 5 projects with Non governmental organizations.<br />
<br />
They are evolving in order to become an institution and in process of building a propper infrastructure that would allow the resolution of high impact issues, as much as bigger workshops, hackatons, talks, conferences and developments. <br />
<br />
You can find out more about this creative and talented group of people at: http://garagelab.tumblr.com/ or their twitter http://twitter.com/garagelab<br />
|<br />
[[File:Bsas2012-Gl.png | 285px]]<br />
|}<br />
<br />
== Learn and play session ==<br />
{|<br />
|<br />
Of the several reunions that we had at Garage Lab, perhaps the most interesting was the one in which one of our members taught Synthetic Biology to a group of computer scientists and engineers through a game. The key question was not how does DNA works but How can we make it work for us?<br />
<br />
Through these talks we find out how people from different disciplines other than natural sciences come up with new ideas and points of view that challenge and make us rethink biology.<br />
<br />
We are planing to develop a game (either a computer game, a cards game or both) for spreading synthetic biology concepts and strategies among young and non specialized people<br />
<br />
| width="30%" align="center" |<br />
{| style="width: 50%"<br />
| align="center" | [http://vimeo.com/45026664 [[File:Bsas2012-Garageee.jpg|220px]]]<br />
|-<br />
| style="text-align: center;" |[http://vimeo.com/45026664 Video]: SynBiology from a simple Computer Scientist point of view.<br />
|}<br />
<br />
|}<br />
<br />
<br />
== Future plans: Solving local problems ==<br />
{|<br />
| width="26%" | [[File:Bsas2012-Riachue.jpg|240px]]<br />
| At the beggining of 2012, Garage Lab contacted our instructors to begin a productive relationship. They believe in the impact the use of Synthetic Biology's standard parts of could have in Argentina, as a tool to help solve endemic issues and they want it to become part of their projects and society. Since then we had a formal presentation of the group and our main project at a Garage Lab Meeting and several of our team members have attended the group meetings often. <br />
<br />
Shortly after the first contact with Garage Lab, they came to our laboratory and together we planned on giving use to the parts that come in the Kit in order to assess a key problem in Buenos Aires, which is the contamination present at Riachuelo, a branch of Río de la Plata. <br />
<br />
Several thousands of people live on the banks and nearby this highly polluted area which has been proven very hard to remediate from the environmental point of view, but also, a political challenge given its social and economic implications. <br />
|}<br />
<br />
{|<br />
|As a iGEM team we are thrilled to be part of this competition, but our ultimate goal is to spread the word about this discipline that it's still ''new'' and ''unexplored'' here, not only to the general public but in the scientific comunnity. Our collaboration with GarageLab will be a sort of testrun;a direct application of the ideas we've comunicated about the Synthetic Biology Community and specifically the possibilities of iGEM developed free technology to help solve both big and small problems in our country. <br />
<br />
Therefore, we decided to explore the question Garage Lab presented us as a secondary project for after the Jamborees, to develop a way to identify and measure water contamination; such as heavy metals and maybe other noxious components at Riachuelo in the form of an unexpensive kit that could be distributed so that each community is able to periodically measure contamination levels in their area. '''The infrastructure is not present... ''''<br />
<br />
We aim to build a set of biosensors using the parts from the Registry, more particulary the arsenic biosensor. We have already contacted Edinburgh 2006, Cambridge 2009 and SouthBend 2011 Teams regarding the procedures and experiences the have had related to arsenic detection using biobricks. We found an amazing support, ideas and encouragement from every team we contacted, including teachers and students.<br />
<br />
<br />
'''So, work will continue after the Jamborees and this is just the start!'''<br />
<br />
| [[File:Bsas2012-manu.jpg|240px]]<br />
|}<br />
<br />
= Impact in the media =<br />
<br />
== After Regional Jamboree ==<br />
<br />
"[http://www.eldia.com.ar/edis/20121022/alumnos-argentinos-primera-vez-mundial-biologia-informaciongeneral1.htm Alumnos argentinos, por primera vez en un “Mundial” de biología]" Diario el Día, 10/22/12.<br />
<br />
"[http://www.diariohoy.net/accion-verNota-id-213948-titulo-Argentinos_clasificaron_para_el_concurso_mundial_de_biolog%C3%ADa_sint%C3%A9tica_ Argentinos clasificaron para el concurso mundial de biología sintética]" Diario Hoy, 10/22/12.<br />
<br />
"[http://www.aptus.com.ar/noticia/2932-competencia-mundial-de-biologia-sintetica Competencia mundial de Biología Sintética]" Aptus Noticias educativas, 10/21/12.<br />
<br />
"[http://www.telam.com.ar/nota/41479/ Argentinos clasificaron para el concurso mundial de biología sintética]" Agencia de noticias TELAM, 10/20/12.<br />
<br />
"[http://www.tvperu.gob.pe/noticias/tecnologia/otros/38160-argentinos-clasificaron-para-el-concurso-mundial-de-biologia-sintetica.html Argentinos clasificaron para el concurso mundial de biología sintética]" TV Perú Noticias , 10/20/12.<br />
<br />
"[http://www.agenciacyta.org.ar/2012/10/debut-y-exito-argentino-en-olimpiadas-de-biologia-sintetica/ Debut y éxito argentino en olimpiadas de biología sintética]" Nota en el portal CyTA. 10/19/12.<br />
<br />
"[http://www.tomamateyavivate.com.ar/formacion-y-universidades-argentinas/debut-y-exito-argentino-en-olimpiadas-de-biologia-sintetica/ Debut y éxito argentino en olimpiadas de biología sintética]" Tomamateyavivate, 10/19/12.<br />
<br />
"[http://www.conicet.gov.ar/?p=2827 Estudiantes argentinos pasan a la etapa final de un certamen mundial]" Nota en la página de noticias del CONICET, 10/19/2012.<br />
<br />
"[http://www.pagina12.com.ar/diario/sociedad/3-206038-2012-10-21.html Elegidos del Conicet]" Página 12, 20/10/12.<br />
<br />
== Before Regional Jamboree ==<br />
<br />
"[http://blogs.scientificamerican.com/lab-rat/2012/09/09/igem-buenos-aires-synthetic-bacterial-communities/ iGEM Buenos Aires: Synthetic bacterial communities]" Lab Rat blog at scientificamerican.com 9/9/12.</div>Lucho.Morohttp://2012.igem.org/Team:Buenos_Aires/HP/GarageLabTeam:Buenos Aires/HP/GarageLab2012-10-27T02:04:13Z<p>Lucho.Moro: /* Future plans: Solving local problems */</p>
<hr />
<div>{{:Team:Buenos_Aires/Templates/menu}}<br />
<br />
= Garage Lab =<br />
{|<br />
|<br />
GarageLab is a group of scientists, artists and engineers cofounded by Emiliano Kargieman and Dario Wainer in 2009 as a Workshop about Bioengineering at the University of Palermo.<br />
<br />
During these years, it grew up to be a multitasking group, by getting involved in subjects such as Neuro-robotics, Contamination assessment, Data Privacy and Management, Personalized Medicine, Public Information and Problematics of Open Innovation.<br />
<br />
Now a days, the group has 22 active members and 5 projects with Non governmental organizations.<br />
<br />
They are evolving in order to become an institution and in process of building a propper infrastructure that would allow the resolution of high impact issues, as much as bigger workshops, hackatons, talks, conferences and developments. <br />
<br />
You can find out more about this creative and talented group of people at: http://garagelab.tumblr.com/ or their twitter http://twitter.com/garagelab<br />
|<br />
[[File:Bsas2012-Gl.png | 285px]]<br />
|}<br />
<br />
== Learn and play session ==<br />
{|<br />
|<br />
Of the several reunions that we had at Garage Lab, perhaps the most interesting was the one in which one of our members taught Synthetic Biology to a group of computer scientists and engineers through a game. The key question was not how does DNA works but How can we make it work for us?<br />
<br />
Through these talks we find out how people from different disciplines other than natural sciences come up with new ideas and points of view that challenge and make us rethink biology.<br />
<br />
We are planing to develop a game (either a computer game, a cards game or both) for spreading synthetic biology concepts and strategies among young and non specialized people<br />
<br />
| width="30%" align="center" |<br />
{| style="width: 50%"<br />
| align="center" | [http://vimeo.com/45026664 [[File:Bsas2012-Garageee.jpg|220px]]]<br />
|-<br />
| style="text-align: center;" |[http://vimeo.com/45026664 Video]: SynBiology from a simple Computer Scientist point of view.<br />
|}<br />
<br />
|}<br />
<br />
<br />
== Future plans: Solving local problems ==<br />
{|<br />
| width="26%" | [[File:Bsas2012-Riachue.jpg|240px]]<br />
| At the beggining of 2012, Garage Lab contacted our instructors to begin a productive relationship. They believe in the impact the use of Synthetic Biology's standard parts of could have in Argentina, as a tool to help solve endemic issues and they want it to become part of their projects and society. Since then we had a formal presentation of the group and our main project at a Garage Lab Meeting and several of our team members have attended the group meetings often. <br />
<br />
Shortly after the first contact with Garage Lab, they came to our laboratory and together we planned on giving use to the parts that come in the Kit in order to assess a key problem in Buenos Aires, which is the contamination present at Riachuelo, a branch of Río de la Plata. <br />
<br />
Several thousands of people live on the banks and nearby this highly polluted area which has been proven very hard to remediate from the environmental point of view, but also, a political challenge given its social and economic implications. <br />
|}<br />
<br />
{|<br />
|As a iGEM team we are thrilled to be part of this competition, but our ultimate goal is to spread the word about this discipline that it's still ''new'' and ''unexplored'' here, not only to the general public but in the scientific comunnity. Our collaboration with GarageLab will be a sort of testrun;a direct application of the ideas we've comunicated about the Synthetic Biology Community and specifically the possibilities of iGEM developed free technology to help solve both big and small problems in our country. <br />
<br />
Therefore, we decided to explore the question Garage Lab presented us as a secondary project for after the Jamborees, to develop a way to identify and measure water contamination; such as heavy metals and maybe other noxious components at Riachuelo in the form of an unexpensive kit that could be distributed so that each community is able to periodically measure contamination levels in their area. '''The infrastructure is not present... ''''<br />
<br />
We aim to build a set of biosensors using the parts from the Registry, more particulary the arsenic biosensor. We have already contacted Edinburgh 2006, Cambridge 2009 and SouthBend 2011 Teams regarding the procedures and experiences the have had related to arsenic detection using biobricks. We found an amazing support, ideas and encouragement from every team we contacted, including teachers and students.<br />
<br />
<br />
'''So, work will continue after the Jamborees and this is just the start!'''<br />
<br />
| [[File:Bsas2012-manu.jpg|240px]]<br />
|}<br />
<br />
= Impact in the media =<br />
<br />
"[http://www.eldia.com.ar/edis/20121022/alumnos-argentinos-primera-vez-mundial-biologia-informaciongeneral1.htm Alumnos argentinos, por primera vez en un “Mundial” de biología]" Diario el Día, 10/22/12.<br />
<br />
"[http://www.diariohoy.net/accion-verNota-id-213948-titulo-Argentinos_clasificaron_para_el_concurso_mundial_de_biolog%C3%ADa_sint%C3%A9tica_ Argentinos clasificaron para el concurso mundial de biología sintética]" Diario Hoy, 10/22/12.<br />
<br />
"[http://www.aptus.com.ar/noticia/2932-competencia-mundial-de-biologia-sintetica Competencia mundial de Biología Sintética]" Aptus Noticias educativas, 10/21/12.<br />
<br />
"[http://www.telam.com.ar/nota/41479/ Argentinos clasificaron para el concurso mundial de biología sintética]" Agencia de noticias TELAM, 10/20/12.<br />
<br />
"[http://www.tvperu.gob.pe/noticias/tecnologia/otros/38160-argentinos-clasificaron-para-el-concurso-mundial-de-biologia-sintetica.html Argentinos clasificaron para el concurso mundial de biología sintética]" TV Perú Noticias , 10/20/12.<br />
<br />
"[http://www.agenciacyta.org.ar/2012/10/debut-y-exito-argentino-en-olimpiadas-de-biologia-sintetica/ Debut y éxito argentino en olimpiadas de biología sintética]" Nota en el portal CyTA. 10/19/12.<br />
<br />
"[http://www.tomamateyavivate.com.ar/formacion-y-universidades-argentinas/debut-y-exito-argentino-en-olimpiadas-de-biologia-sintetica/ Debut y éxito argentino en olimpiadas de biología sintética]" Tomamateyavivate, 10/19/12.<br />
<br />
"[http://www.conicet.gov.ar/?p=2827 Estudiantes argentinos pasan a la etapa final de un certamen mundial]" Nota en la página de noticias del CONICET, 10/19/2012.<br />
<br />
"[http://www.pagina12.com.ar/diario/sociedad/3-206038-2012-10-21.html Elegidos del Conicet]" Página 12, 20/10/12.<br />
<br />
"[http://blogs.scientificamerican.com/lab-rat/2012/09/09/igem-buenos-aires-synthetic-bacterial-communities/ iGEM Buenos Aires: Synthetic bacterial communities]" Lab Rat blog at scientificamerican.com 9/9/12.</div>Lucho.Morohttp://2012.igem.org/Team:Buenos_Aires/HP/GarageLabTeam:Buenos Aires/HP/GarageLab2012-10-27T02:03:11Z<p>Lucho.Moro: /* Impact in the media */</p>
<hr />
<div>{{:Team:Buenos_Aires/Templates/menu}}<br />
<br />
= Garage Lab =<br />
{|<br />
|<br />
GarageLab is a group of scientists, artists and engineers cofounded by Emiliano Kargieman and Dario Wainer in 2009 as a Workshop about Bioengineering at the University of Palermo.<br />
<br />
During these years, it grew up to be a multitasking group, by getting involved in subjects such as Neuro-robotics, Contamination assessment, Data Privacy and Management, Personalized Medicine, Public Information and Problematics of Open Innovation.<br />
<br />
Now a days, the group has 22 active members and 5 projects with Non governmental organizations.<br />
<br />
They are evolving in order to become an institution and in process of building a propper infrastructure that would allow the resolution of high impact issues, as much as bigger workshops, hackatons, talks, conferences and developments. <br />
<br />
You can find out more about this creative and talented group of people at: http://garagelab.tumblr.com/ or their twitter http://twitter.com/garagelab<br />
|<br />
[[File:Bsas2012-Gl.png | 285px]]<br />
|}<br />
<br />
== Learn and play session ==<br />
{|<br />
|<br />
Of the several reunions that we had at Garage Lab, perhaps the most interesting was the one in which one of our members taught Synthetic Biology to a group of computer scientists and engineers through a game. The key question was not how does DNA works but How can we make it work for us?<br />
<br />
Through these talks we find out how people from different disciplines other than natural sciences come up with new ideas and points of view that challenge and make us rethink biology.<br />
<br />
We are planing to develop a game (either a computer game, a cards game or both) for spreading synthetic biology concepts and strategies among young and non specialized people<br />
<br />
| width="30%" align="center" |<br />
{| style="width: 50%"<br />
| align="center" | [http://vimeo.com/45026664 [[File:Bsas2012-Garageee.jpg|220px]]]<br />
|-<br />
| style="text-align: center;" |[http://vimeo.com/45026664 Video]: SynBiology from a simple Computer Scientist point of view.<br />
|}<br />
<br />
|}<br />
<br />
<br />
== Future plans: Solving local problems ==<br />
{|<br />
| width="26%" | [[File:Bsas2012-Riachue.jpg|240px]]<br />
| At the beggining of 2012, Garage Lab contacted our instructors to begin a productive relationship. They believe in the impact the use of Synthetic Biology's standard parts of could have in Argentina, as a tool to help solve endemic issues and they want it to become part of their projects and society. Since then we had a formal presentation of the group and our main project at a Garage Lab Meeting and several of our team members have attended the group meetings often. <br />
<br />
Shortly after the first contact with Garage Lab, they came to our laboratory and together we planned on giving use to the parts that come in the Kit in order to assess a key problem in Buenos Aires, which is the contamination present at Riachuelo, a branch of Río de la Plata. <br />
<br />
Several thousands of people live on the banks and nearby this highly polluted area which has been proven very hard to remediate from the environmental point of view, but also, a political challenge given its social and economic implications. <br />
|}<br />
<br />
{|<br />
|As a iGEM team we are thrilled to be part of this competition, but our ultimate goal is to spread the word about this discipline that it's still ''new'' and ''unexplored'' here, not only to the general public but in the scientific comunnity. Since our participation in the LatinAmerican Jamboree papers!![link]. Our collaboration with GarageLab will be a sort of testrun;a direct application of the ideas we've comunicated about the Synthetic Biology Community and specifically the possibilities of iGEM developed free technology to help solve both big and small problems in our country. <br />
<br />
Therefore, we decided to explore the question Garage Lab presented us as a secondary project for after the Jamborees, to develop a way to identify and measure water contamination; such as heavy metals and maybe other noxious components at Riachuelo in the form of an unexpensive kit that could be distributed so that each community is able to periodically measure contamination levels in their area. '''The infrastructure is not present... ''''<br />
<br />
We aim to build a set of biosensors using the parts from the Registry, more particulary the arsenic biosensor. We have already contacted Edinburgh 2006, Cambridge 2009 and SouthBend 2011 Teams regarding the procedures and experiences the have had related to arsenic detection using biobricks. We found an amazing support, ideas and encouragement from every team we contacted, including teachers and students.<br />
<br />
<br />
'''So, work will continue after the Jamborees and this is just the start!'''<br />
<br />
| [[File:Bsas2012-manu.jpg|240px]]<br />
|}<br />
<br />
<br />
= Impact in the media =<br />
<br />
"[http://www.eldia.com.ar/edis/20121022/alumnos-argentinos-primera-vez-mundial-biologia-informaciongeneral1.htm Alumnos argentinos, por primera vez en un “Mundial” de biología]" Diario el Día, 10/22/12.<br />
<br />
"[http://www.diariohoy.net/accion-verNota-id-213948-titulo-Argentinos_clasificaron_para_el_concurso_mundial_de_biolog%C3%ADa_sint%C3%A9tica_ Argentinos clasificaron para el concurso mundial de biología sintética]" Diario Hoy, 10/22/12.<br />
<br />
"[http://www.aptus.com.ar/noticia/2932-competencia-mundial-de-biologia-sintetica Competencia mundial de Biología Sintética]" Aptus Noticias educativas, 10/21/12.<br />
<br />
"[http://www.telam.com.ar/nota/41479/ Argentinos clasificaron para el concurso mundial de biología sintética]" Agencia de noticias TELAM, 10/20/12.<br />
<br />
"[http://www.tvperu.gob.pe/noticias/tecnologia/otros/38160-argentinos-clasificaron-para-el-concurso-mundial-de-biologia-sintetica.html Argentinos clasificaron para el concurso mundial de biología sintética]" TV Perú Noticias , 10/20/12.<br />
<br />
"[http://www.agenciacyta.org.ar/2012/10/debut-y-exito-argentino-en-olimpiadas-de-biologia-sintetica/ Debut y éxito argentino en olimpiadas de biología sintética]" Nota en el portal CyTA. 10/19/12.<br />
<br />
"[http://www.tomamateyavivate.com.ar/formacion-y-universidades-argentinas/debut-y-exito-argentino-en-olimpiadas-de-biologia-sintetica/ Debut y éxito argentino en olimpiadas de biología sintética]" Tomamateyavivate, 10/19/12.<br />
<br />
"[http://www.conicet.gov.ar/?p=2827 Estudiantes argentinos pasan a la etapa final de un certamen mundial]" Nota en la página de noticias del CONICET, 10/19/2012.<br />
<br />
"[http://www.pagina12.com.ar/diario/sociedad/3-206038-2012-10-21.html Elegidos del Conicet]" Página 12, 20/10/12.<br />
<br />
"[http://blogs.scientificamerican.com/lab-rat/2012/09/09/igem-buenos-aires-synthetic-bacterial-communities/ iGEM Buenos Aires: Synthetic bacterial communities]" Lab Rat blog at scientificamerican.com 9/9/12.</div>Lucho.Morohttp://2012.igem.org/Team:Buenos_Aires/HP/GarageLabTeam:Buenos Aires/HP/GarageLab2012-10-27T02:02:29Z<p>Lucho.Moro: /* Impact in the media */</p>
<hr />
<div>{{:Team:Buenos_Aires/Templates/menu}}<br />
<br />
= Garage Lab =<br />
{|<br />
|<br />
GarageLab is a group of scientists, artists and engineers cofounded by Emiliano Kargieman and Dario Wainer in 2009 as a Workshop about Bioengineering at the University of Palermo.<br />
<br />
During these years, it grew up to be a multitasking group, by getting involved in subjects such as Neuro-robotics, Contamination assessment, Data Privacy and Management, Personalized Medicine, Public Information and Problematics of Open Innovation.<br />
<br />
Now a days, the group has 22 active members and 5 projects with Non governmental organizations.<br />
<br />
They are evolving in order to become an institution and in process of building a propper infrastructure that would allow the resolution of high impact issues, as much as bigger workshops, hackatons, talks, conferences and developments. <br />
<br />
You can find out more about this creative and talented group of people at: http://garagelab.tumblr.com/ or their twitter http://twitter.com/garagelab<br />
|<br />
[[File:Bsas2012-Gl.png | 285px]]<br />
|}<br />
<br />
== Learn and play session ==<br />
{|<br />
|<br />
Of the several reunions that we had at Garage Lab, perhaps the most interesting was the one in which one of our members taught Synthetic Biology to a group of computer scientists and engineers through a game. The key question was not how does DNA works but How can we make it work for us?<br />
<br />
Through these talks we find out how people from different disciplines other than natural sciences come up with new ideas and points of view that challenge and make us rethink biology.<br />
<br />
We are planing to develop a game (either a computer game, a cards game or both) for spreading synthetic biology concepts and strategies among young and non specialized people<br />
<br />
| width="30%" align="center" |<br />
{| style="width: 50%"<br />
| align="center" | [http://vimeo.com/45026664 [[File:Bsas2012-Garageee.jpg|220px]]]<br />
|-<br />
| style="text-align: center;" |[http://vimeo.com/45026664 Video]: SynBiology from a simple Computer Scientist point of view.<br />
|}<br />
<br />
|}<br />
<br />
<br />
== Future plans: Solving local problems ==<br />
{|<br />
| width="26%" | [[File:Bsas2012-Riachue.jpg|240px]]<br />
| At the beggining of 2012, Garage Lab contacted our instructors to begin a productive relationship. They believe in the impact the use of Synthetic Biology's standard parts of could have in Argentina, as a tool to help solve endemic issues and they want it to become part of their projects and society. Since then we had a formal presentation of the group and our main project at a Garage Lab Meeting and several of our team members have attended the group meetings often. <br />
<br />
Shortly after the first contact with Garage Lab, they came to our laboratory and together we planned on giving use to the parts that come in the Kit in order to assess a key problem in Buenos Aires, which is the contamination present at Riachuelo, a branch of Río de la Plata. <br />
<br />
Several thousands of people live on the banks and nearby this highly polluted area which has been proven very hard to remediate from the environmental point of view, but also, a political challenge given its social and economic implications. <br />
|}<br />
<br />
{|<br />
|As a iGEM team we are thrilled to be part of this competition, but our ultimate goal is to spread the word about this discipline that it's still ''new'' and ''unexplored'' here, not only to the general public but in the scientific comunnity. Since our participation in the LatinAmerican Jamboree papers!![link]. Our collaboration with GarageLab will be a sort of testrun;a direct application of the ideas we've comunicated about the Synthetic Biology Community and specifically the possibilities of iGEM developed free technology to help solve both big and small problems in our country. <br />
<br />
Therefore, we decided to explore the question Garage Lab presented us as a secondary project for after the Jamborees, to develop a way to identify and measure water contamination; such as heavy metals and maybe other noxious components at Riachuelo in the form of an unexpensive kit that could be distributed so that each community is able to periodically measure contamination levels in their area. '''The infrastructure is not present... ''''<br />
<br />
We aim to build a set of biosensors using the parts from the Registry, more particulary the arsenic biosensor. We have already contacted Edinburgh 2006, Cambridge 2009 and SouthBend 2011 Teams regarding the procedures and experiences the have had related to arsenic detection using biobricks. We found an amazing support, ideas and encouragement from every team we contacted, including teachers and students.<br />
<br />
<br />
'''So, work will continue after the Jamborees and this is just the start!'''<br />
<br />
| [[File:Bsas2012-manu.jpg|240px]]<br />
|}<br />
<br />
<br />
= Impact in the media =<br />
<br />
"[http://www.eldia.com.ar/edis/20121022/alumnos-argentinos-primera-vez-mundial-biologia-informaciongeneral1.htm Alumnos argentinos, por primera vez en un “Mundial” de biología]" Diario el Día, 10/22/12.<br />
<br />
"[http://www.diariohoy.net/accion-verNota-id-213948-titulo-Argentinos_clasificaron_para_el_concurso_mundial_de_biolog%C3%ADa_sint%C3%A9tica_ Argentinos clasificaron para el concurso mundial de biología sintética]" Diario Hoy, 10/22/12.<br />
<br />
"[http://www.aptus.com.ar/noticia/2932-competencia-mundial-de-biologia-sintetica Competencia mundial de Biología Sintética]" Aptus Noticias educativas, 10/21/12.<br />
<br />
"[http://www.telam.com.ar/nota/41479/ Argentinos clasificaron para el concurso mundial de biología sintética]" Agencia de noticias TELAM, 10/20/12.<br />
<br />
"[http://www.tvperu.gob.pe/noticias/tecnologia/otros/38160-argentinos-clasificaron-para-el-concurso-mundial-de-biologia-sintetica.html Argentinos clasificaron para el concurso mundial de biología sintética]" TV Perú Noticias , 10/20/12.<br />
<br />
"[http://www.agenciacyta.org.ar/2012/10/debut-y-exito-argentino-en-olimpiadas-de-biologia-sintetica/ Debut y éxito argentino en olimpiadas de biología sintética]" Nota en el portal CyTA. 10/19/12.<br />
<br />
"[http://www.tomamateyavivate.com.ar/formacion-y-universidades-argentinas/debut-y-exito-argentino-en-olimpiadas-de-biologia-sintetica/ Debut y éxito argentino en olimpiadas de biología sintética]" Tomamateyavivate, 10/19/12.<br />
<br />
"[http://www.conicet.gov.ar/?p=2827 Estudiantes argentinos pasan a la etapa final de un certamen mundial]" Nota en la página de noticias del CONICET, 10/19/2012.<br />
<br />
"[http://www.pagina12.com.ar/diario/sociedad/3-206038-2012-10-21.html Elegidos del Conicet]" Página 12, 20/10/12.<br />
"[http://blogs.scientificamerican.com/lab-rat/2012/09/09/igem-buenos-aires-synthetic-bacterial-communities/ iGEM Buenos Aires: Synthetic bacterial communities]" Lab Rat blog at scientificamerican.com 9/9/12.</div>Lucho.Morohttp://2012.igem.org/Team:Buenos_Aires/HP/GarageLabTeam:Buenos Aires/HP/GarageLab2012-10-27T01:58:48Z<p>Lucho.Moro: /* Impact in the media */</p>
<hr />
<div>{{:Team:Buenos_Aires/Templates/menu}}<br />
<br />
= Garage Lab =<br />
{|<br />
|<br />
GarageLab is a group of scientists, artists and engineers cofounded by Emiliano Kargieman and Dario Wainer in 2009 as a Workshop about Bioengineering at the University of Palermo.<br />
<br />
During these years, it grew up to be a multitasking group, by getting involved in subjects such as Neuro-robotics, Contamination assessment, Data Privacy and Management, Personalized Medicine, Public Information and Problematics of Open Innovation.<br />
<br />
Now a days, the group has 22 active members and 5 projects with Non governmental organizations.<br />
<br />
They are evolving in order to become an institution and in process of building a propper infrastructure that would allow the resolution of high impact issues, as much as bigger workshops, hackatons, talks, conferences and developments. <br />
<br />
You can find out more about this creative and talented group of people at: http://garagelab.tumblr.com/ or their twitter http://twitter.com/garagelab<br />
|<br />
[[File:Bsas2012-Gl.png | 285px]]<br />
|}<br />
<br />
== Learn and play session ==<br />
{|<br />
|<br />
Of the several reunions that we had at Garage Lab, perhaps the most interesting was the one in which one of our members taught Synthetic Biology to a group of computer scientists and engineers through a game. The key question was not how does DNA works but How can we make it work for us?<br />
<br />
Through these talks we find out how people from different disciplines other than natural sciences come up with new ideas and points of view that challenge and make us rethink biology.<br />
<br />
We are planing to develop a game (either a computer game, a cards game or both) for spreading synthetic biology concepts and strategies among young and non specialized people<br />
<br />
| width="30%" align="center" |<br />
{| style="width: 50%"<br />
| align="center" | [http://vimeo.com/45026664 [[File:Bsas2012-Garageee.jpg|220px]]]<br />
|-<br />
| style="text-align: center;" |[http://vimeo.com/45026664 Video]: SynBiology from a simple Computer Scientist point of view.<br />
|}<br />
<br />
|}<br />
<br />
<br />
== Future plans: Solving local problems ==<br />
{|<br />
| width="26%" | [[File:Bsas2012-Riachue.jpg|240px]]<br />
| At the beggining of 2012, Garage Lab contacted our instructors to begin a productive relationship. They believe in the impact the use of Synthetic Biology's standard parts of could have in Argentina, as a tool to help solve endemic issues and they want it to become part of their projects and society. Since then we had a formal presentation of the group and our main project at a Garage Lab Meeting and several of our team members have attended the group meetings often. <br />
<br />
Shortly after the first contact with Garage Lab, they came to our laboratory and together we planned on giving use to the parts that come in the Kit in order to assess a key problem in Buenos Aires, which is the contamination present at Riachuelo, a branch of Río de la Plata. <br />
<br />
Several thousands of people live on the banks and nearby this highly polluted area which has been proven very hard to remediate from the environmental point of view, but also, a political challenge given its social and economic implications. <br />
|}<br />
<br />
{|<br />
|As a iGEM team we are thrilled to be part of this competition, but our ultimate goal is to spread the word about this discipline that it's still ''new'' and ''unexplored'' here, not only to the general public but in the scientific comunnity. Since our participation in the LatinAmerican Jamboree papers!![link]. Our collaboration with GarageLab will be a sort of testrun;a direct application of the ideas we've comunicated about the Synthetic Biology Community and specifically the possibilities of iGEM developed free technology to help solve both big and small problems in our country. <br />
<br />
Therefore, we decided to explore the question Garage Lab presented us as a secondary project for after the Jamborees, to develop a way to identify and measure water contamination; such as heavy metals and maybe other noxious components at Riachuelo in the form of an unexpensive kit that could be distributed so that each community is able to periodically measure contamination levels in their area. '''The infrastructure is not present... ''''<br />
<br />
We aim to build a set of biosensors using the parts from the Registry, more particulary the arsenic biosensor. We have already contacted Edinburgh 2006, Cambridge 2009 and SouthBend 2011 Teams regarding the procedures and experiences the have had related to arsenic detection using biobricks. We found an amazing support, ideas and encouragement from every team we contacted, including teachers and students.<br />
<br />
<br />
'''So, work will continue after the Jamborees and this is just the start!'''<br />
<br />
| [[File:Bsas2012-manu.jpg|240px]]<br />
|}<br />
<br />
<br />
= Impact in the media =<br />
<br />
"[http://www.eldia.com.ar/edis/20121022/alumnos-argentinos-primera-vez-mundial-biologia-informaciongeneral1.htm Alumnos argentinos, por primera vez en un “Mundial” de biología]" Diario el Día, 22/10/12.<br />
<br />
"[http://www.diariohoy.net/accion-verNota-id-213948-titulo-Argentinos_clasificaron_para_el_concurso_mundial_de_biolog%C3%ADa_sint%C3%A9tica_ Argentinos clasificaron para el concurso mundial de biología sintética]" Diario Hoy, 22/10/12.<br />
<br />
"[http://www.telam.com.ar/nota/41479/ Argentinos clasificaron para el concurso mundial de biología sintética]" Agencia de noticias TELAM, 20/10/12.<br />
<br />
"[http://www.agenciacyta.org.ar/2012/10/debut-y-exito-argentino-en-olimpiadas-de-biologia-sintetica/ Debut y éxito argentino en olimpiadas de biología sintética]" Nota en el portal CyTA. 19/10/12.<br />
<br />
"[http://www.tomamateyavivate.com.ar/formacion-y-universidades-argentinas/debut-y-exito-argentino-en-olimpiadas-de-biologia-sintetica/ Debut y éxito argentino en olimpiadas de biología sintética]" Tomamateyavivate, 19/10/12.<br />
<br />
"[http://www.aptus.com.ar/noticia/2932-competencia-mundial-de-biologia-sintetica Competencia mundial de Biología Sintética]" Aptus Noticias educativas, 21/10/12.<br />
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"[http://www.tvperu.gob.pe/noticias/tecnologia/otros/38160-argentinos-clasificaron-para-el-concurso-mundial-de-biologia-sintetica.html Argentinos clasificaron para el concurso mundial de biología sintética]" TV Perú Noticias , 20/10/12.<br />
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"[http://www.conicet.gov.ar/?p=2827 Estudiantes argentinos pasan a la etapa final de un certamen mundial]" Nota en la página de noticias del CONICET, 19/10/2012.<br />
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"[http://www.pagina12.com.ar/diario/sociedad/3-206038-2012-10-21.html Elegidos del Conicet]" Página 12, 20/10/12.<br />
"[http://blogs.scientificamerican.com/lab-rat/2012/09/09/igem-buenos-aires-synthetic-bacterial-communities/ iGEM Buenos Aires: Synthetic bacterial communities]" Lab Rat blog at scientificamerican.com 9/9/12.</div>Lucho.Morohttp://2012.igem.org/Team:Buenos_Aires/HP/GarageLabTeam:Buenos Aires/HP/GarageLab2012-10-27T01:58:12Z<p>Lucho.Moro: /* Garage Lab */</p>
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= Garage Lab =<br />
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GarageLab is a group of scientists, artists and engineers cofounded by Emiliano Kargieman and Dario Wainer in 2009 as a Workshop about Bioengineering at the University of Palermo.<br />
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During these years, it grew up to be a multitasking group, by getting involved in subjects such as Neuro-robotics, Contamination assessment, Data Privacy and Management, Personalized Medicine, Public Information and Problematics of Open Innovation.<br />
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Now a days, the group has 22 active members and 5 projects with Non governmental organizations.<br />
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They are evolving in order to become an institution and in process of building a propper infrastructure that would allow the resolution of high impact issues, as much as bigger workshops, hackatons, talks, conferences and developments. <br />
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You can find out more about this creative and talented group of people at: http://garagelab.tumblr.com/ or their twitter http://twitter.com/garagelab<br />
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[[File:Bsas2012-Gl.png | 285px]]<br />
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== Learn and play session ==<br />
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Of the several reunions that we had at Garage Lab, perhaps the most interesting was the one in which one of our members taught Synthetic Biology to a group of computer scientists and engineers through a game. The key question was not how does DNA works but How can we make it work for us?<br />
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Through these talks we find out how people from different disciplines other than natural sciences come up with new ideas and points of view that challenge and make us rethink biology.<br />
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We are planing to develop a game (either a computer game, a cards game or both) for spreading synthetic biology concepts and strategies among young and non specialized people<br />
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| align="center" | [http://vimeo.com/45026664 [[File:Bsas2012-Garageee.jpg|220px]]]<br />
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| style="text-align: center;" |[http://vimeo.com/45026664 Video]: SynBiology from a simple Computer Scientist point of view.<br />
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== Future plans: Solving local problems ==<br />
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| At the beggining of 2012, Garage Lab contacted our instructors to begin a productive relationship. They believe in the impact the use of Synthetic Biology's standard parts of could have in Argentina, as a tool to help solve endemic issues and they want it to become part of their projects and society. Since then we had a formal presentation of the group and our main project at a Garage Lab Meeting and several of our team members have attended the group meetings often. <br />
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Shortly after the first contact with Garage Lab, they came to our laboratory and together we planned on giving use to the parts that come in the Kit in order to assess a key problem in Buenos Aires, which is the contamination present at Riachuelo, a branch of Río de la Plata. <br />
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Several thousands of people live on the banks and nearby this highly polluted area which has been proven very hard to remediate from the environmental point of view, but also, a political challenge given its social and economic implications. <br />
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|As a iGEM team we are thrilled to be part of this competition, but our ultimate goal is to spread the word about this discipline that it's still ''new'' and ''unexplored'' here, not only to the general public but in the scientific comunnity. Since our participation in the LatinAmerican Jamboree papers!![link]. Our collaboration with GarageLab will be a sort of testrun;a direct application of the ideas we've comunicated about the Synthetic Biology Community and specifically the possibilities of iGEM developed free technology to help solve both big and small problems in our country. <br />
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Therefore, we decided to explore the question Garage Lab presented us as a secondary project for after the Jamborees, to develop a way to identify and measure water contamination; such as heavy metals and maybe other noxious components at Riachuelo in the form of an unexpensive kit that could be distributed so that each community is able to periodically measure contamination levels in their area. '''The infrastructure is not present... ''''<br />
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We aim to build a set of biosensors using the parts from the Registry, more particulary the arsenic biosensor. We have already contacted Edinburgh 2006, Cambridge 2009 and SouthBend 2011 Teams regarding the procedures and experiences the have had related to arsenic detection using biobricks. We found an amazing support, ideas and encouragement from every team we contacted, including teachers and students.<br />
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'''So, work will continue after the Jamborees and this is just the start!'''<br />
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= Impact in the media =<br />
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"[http://www.eldia.com.ar/edis/20121022/alumnos-argentinos-primera-vez-mundial-biologia-informaciongeneral1.htm Alumnos argentinos, por primera vez en un “Mundial” de biología]" Diario el Día, 22/10/12.<br />
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"[http://www.diariohoy.net/accion-verNota-id-213948-titulo-Argentinos_clasificaron_para_el_concurso_mundial_de_biolog%C3%ADa_sint%C3%A9tica_ Argentinos clasificaron para el concurso mundial de biología sintética]" Diario Hoy, 22/10/12.<br />
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"[http://www.telam.com.ar/nota/41479/ Argentinos clasificaron para el concurso mundial de biología sintética]" Agencia de noticias TELAM, 20/10/12.<br />
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"[http://www.agenciacyta.org.ar/2012/10/debut-y-exito-argentino-en-olimpiadas-de-biologia-sintetica/ Debut y éxito argentino en olimpiadas de biología sintética]" Nota en el portal CyTA. 19/10/12.<br />
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"[http://www.tomamateyavivate.com.ar/formacion-y-universidades-argentinas/debut-y-exito-argentino-en-olimpiadas-de-biologia-sintetica/ Debut y éxito argentino en olimpiadas de biología sintética]" Tomamateyavivate, 19/10/12.<br />
"[http://www.aptus.com.ar/noticia/2932-competencia-mundial-de-biologia-sintetica Competencia mundial de Biología Sintética]" Aptus Noticias educativas, 21/10/12.<br />
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"[http://www.tvperu.gob.pe/noticias/tecnologia/otros/38160-argentinos-clasificaron-para-el-concurso-mundial-de-biologia-sintetica.html Argentinos clasificaron para el concurso mundial de biología sintética]" TV Perú Noticias , 20/10/12.<br />
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"[http://www.conicet.gov.ar/?p=2827 Estudiantes argentinos pasan a la etapa final de un certamen mundial]" Nota en la página de noticias del CONICET, 19/10/2012.<br />
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"[http://www.pagina12.com.ar/diario/sociedad/3-206038-2012-10-21.html Elegidos del Conicet]" Página 12, 20/10/12.<br />
"[http://blogs.scientificamerican.com/lab-rat/2012/09/09/igem-buenos-aires-synthetic-bacterial-communities/ iGEM Buenos Aires: Synthetic bacterial communities]" Lab Rat blog at scientificamerican.com 9/9/12.</div>Lucho.Moro