Team:Lyon-INSA/notebook

From 2012.igem.org

(Difference between revisions)
 
(19 intermediate revisions not shown)
Line 19: Line 19:
$(document).ready(function(){
$(document).ready(function(){
 +
$("#lab").click();
$("#noteBookTextContent").tinyscrollbar();
$("#noteBookTextContent").tinyscrollbar();
Line 342: Line 343:
<body>
<body>
-
 
-
 
-
<div id="projectBar" class="menuBar">
 
-
    <div class="boutonMenu" onclick="window.location='menu';">Menu</div>
 
-
    <div id="ongletBar">
 
-
        <div class="boutonOnglet"  onclick="window.location='project';">Project</div>
 
-
        <div class="boutonOnglet"  onclick="window.location='modelling';">Modelling</div>
 
-
        <div class="boutonOnglet"  onclick="window.location='safety';">Safety</div>
 
-
        <div class="boutonOnglet BOClicked">Notebook</div>
 
-
        <div class="boutonOnglet" onclick="window.location='protocol';">Protocols</div>
 
-
        <div class="boutonOnglet"  onclick="window.location='datapage';">Data Page</div>
 
-
        <div class="boutonOnglet"  onclick="window.location='results';">Results</div>
 
-
        <div class="boutonOnglet" onclick="window.location='Achievements';">Achievements</div>
 
-
    </div>
 
-
</div>
 
-
<div id="bandeau"></div>
 
<div id="xml" style="display:none;">
<div id="xml" style="display:none;">
Line 2,233: Line 2,218:
</jour>
</jour>
 +
</month>
 +
<month name="October" size="31">
 +
<jour nb="10">
 +
                        <date>Wednesday, October 10th 2012</date>
 +
 +
<titre>Surfactant</titre>
 +
<description>
 +
<ul>
 +
<li>Ligation of the part pBK14 containing the <i>gfp</i> gene with the part pBK7 (containing a <i>B. subtilis</i> RBS).</li>
 +
<li>Ligation of the part pBK13 containing the <i>rfp</i> gene with the part pBK7 (containing a <i>B. subtilis</i> RBS).</li>
 +
<li>Transformation of the NM522 <i>E. coli</i> strain with the two ligations.</li>
 +
</ul>
 +
</description>
 +
<titre>Stick</titre>
 +
<description>Ligation of the PCR product <i>xylR</i> gene with the part pBK9 (P<sub><i>lac</i></sub> gene)and NM522 transformation.
 +
</description>
 +
</jour>
 +
<jour nb="11">
 +
<date>Thursday, October 11th 2012</date>
 +
 +
<titre>Surfactant</titre>
 +
<description>
 +
Screening of 15 clones per transformation (antibiotic resistance test).
 +
</description>
 +
<titre>Stick</titre>
 +
<description>
 +
Inoculation of 5 mL of LB medium with chloramphenicol with 1 colony of <i>Bacillus subtilis</i> 168 <i>ΔabrB</i> and 5 mL of LB medium with 1 colony of <i>Bacillus subtilis</i> 168. Incubation overnight at 37°C. (This is for the 48h positive <i>Bacillus subtilis</i> biofilm test plate)
 +
</description>
 +
</jour>
 +
<jour nb="12">
 +
<date>Friday, October 12th 2012</date>
 +
 +
<titre>Surfactant</titre>
 +
<description>
 +
Liquid cultures are inoculated with colonies having the expected antibiotic phenotype.
 +
</br>
 +
To test the surfactant properties of the surfactin produced by the <i>B. subtilis</i> BK52 strain, a biofilm test was made in a 24-well plate. Liquid cultures are seeded with BK52 and BK49.
 +
</description>
 +
<titre>Stick</titre>
 +
<description>
 +
<ul>
 +
<li>(24h plate)The same inoculation as previously is prepared.</li>
 +
<li>(48h plate)<i>Bacillus subtilis</i> 168 <i>ΔabrB</i> or <i>Bacillus subtilis</i> 168 are added in the 12 well plate with MgSO4  1mM and Glucose 0.1 %.</li>
 +
</ul>
 +
The plate is incubated at 37°C for 48h
-
</month>
+
(For both) Inoculation of 5 mL of LB medium with 1 colony of <i>E coli</i> (<i>ompR++</i> GFP, curlis overproduction). Incubation overnight at 37°C.
 +
</description>
 +
</jour>
 +
<jour nb="13">
 +
<date>Saturday, October 13th 2012</date>
 +
 +
<titre>Surfactant</titre>
 +
<description>
 +
Miniprep of the screened clones. Unfortunately, the gel electrophoresis was disappointing and the extracted plasmids did not contain the ligated genes.
 +
</br>
 +
A 24-well plate is incubated with filtered supernatant of the saturated cultures with BK52 and BK49 at room temperature for 24 hours.
 +
 +
</description>
 +
<titre>Stick</titre>
 +
<description>
 +
(24h plate)<i>Bacillus subtilis</i> 168 <i>ΔabrB</i> or <i>Bacillus subtilis</i> 168 are added in a 12-well plate with MgSO4  1mM and Glucose 0.1 %.
 +
The plate is incubated at 37°C for 48h.
 +
</description>
 +
</jour>
 +
 +
<jour nb="14">
 +
<date>Sunday, October 14th 2012</date>
 +
 +
<titre>Surfactant</titre>
 +
<description>
 +
The wells are carefully rinsed with M63 medium two times after having previously discarded the supernatant. Then, each well is inoculated with a saturated adherent <i>E. coli</i> strain diluted 100 times in LB medium diluted 2 times.</br>
 +
Moreover, another 24-well plate assay was made using the same supernatant, but this time the plate was incubated only 4 hours before <i>E. coli</i> inoculation.
 +
 +
</description>
 +
<titre>Stick</titre>
 +
<description>
 +
(For both) The supernatant is removed, wells are washed and LB is added.To see if <i>E coli</i> forms a biofilm over the <i>Bacillus subtilis</i> biofilm, the overnight culture of <i>E coli</i> is added into each well.
 +
Plates are incubated for 36h at 30°C.
 +
</description>
 +
</jour>
 +
 +
<jour nb="16">
 +
<date>Tuesday, October 16th 2012</date>
 +
 +
<titre>Surfactant</titre>
 +
<description>
 +
Unfortunately, the observation of the plates under confocal microscope is not possible, so the experiment must be repeated. The only difference is that this time 12-well plates  with glass lamellae are used so the observation of the biofilm would be possible.
 +
 +
</description>
 +
<titre>Stick</titre>
 +
<description>
 +
(For both) Now let's observe using the confocal microscope !
 +
Results : It works ! Our positive <i>Bacillus subtilis</i> biofilm inhibits the stick of other bacteria !
 +
</description>
 +
</jour>
 +
<jour nb="17">
 +
<date>Wednesday, October 17th 2012</date>
 +
 +
<titre>Surfactant</titre>
 +
<description>
 +
Liquid cultures were inoculated with the strains BK49 and BK52 and then incubated at 37C for 48 hours.
 +
</description>
 +
 +
</jour>
 +
 +
 +
<jour nb="19">
 +
<date>Saturday, October 19th 2012</date>
 +
 +
<titre>Surfactant</titre>
 +
<description>
 +
The supernatant from the plate is discarded. Each well is inoculated with an <i>E. coli</i> saturated culture diluted 50 times in LB medium diluted 2 times.
 +
 +
 +
</description>
 +
 +
</jour>
 +
 +
<jour nb="20">
 +
<date>Sunday, October 20th 2012</date>
 +
 +
<titre>Surfactant</titre>
 +
<description>
 +
Ligation of pBK40 and pBK16 followed by transformation into the NM522 strain.
 +
 +
</description>
 +
 +
</jour>
 +
 +
<jour nb="21">
 +
<date>Monday, October 21st 2012</date>
 +
 +
<titre>Surfactant</titre>
 +
<description>
 +
The surfactin pre-treated glass lamellae are observed under the confocal microscope. The negative control has a thick biofilm whereas the treated lamella has a few isolated colonies.
 +
 +
</description>
 +
 +
</jour>
 +
 +
<jour nb="22">
 +
<date>Tuesday, October 22nd 2012</date>
 +
 +
<titre>Surfactant</titre>
 +
<description>
 +
Screening of 12 clones.
 +
 +
</description>
 +
 +
</jour>
 +
 +
<jour nb="23">
 +
<date>Wednesday, October 23rd 2012</date>
 +
 +
<titre>Surfactant</titre>
 +
<description>
 +
Miniprep of the transformed clones. The gel electrophoresis confirmed the expected insert. (<i>abrB</i> gene in pSB1C3 backbone)
 +
 +
</description>
 +
 +
</jour>
 +
 +
<jour nb="24">
 +
<date>Tuesday, October 24th 2012</date>
 +
 +
<titre>Surfactant</titre>
 +
<description>
 +
The plasmids pBK42 and pBK46 were submitted to the Registry.
 +
 +
</description>
 +
 +
</jour>
 +
 +
</month>
</project>
</project>
Line 2,246: Line 2,404:
</div>
</div>
-
<div id="contentPage">
+
<div id="pageContent">
-
<br/>
+
 
<!--[if IE]>
<!--[if IE]>
     <center><span style="font-size:23px;text-decoration:blink;color:red;">Sorry !! Our notebook doesn't work on IE, please use another browser to enjoy it</span></center>
     <center><span style="font-size:23px;text-decoration:blink;color:red;">Sorry !! Our notebook doesn't work on IE, please use another browser to enjoy it</span></center>
<![endif]-->
<![endif]-->
-
<br/>
 
<h1>The Notebook</h1>
<h1>The Notebook</h1>
<br/>
<br/>
Line 2,258: Line 2,415:
        <div class="month" id="August">August</div>
        <div class="month" id="August">August</div>
                         <div class="month" id="September">September</div>
                         <div class="month" id="September">September</div>
 +
                        <div class="month" id="October">October</div>
 +
</div>
</div>
<br/>
<br/>

Latest revision as of 00:15, 27 October 2012

The Notebook


July
August
September
October


Previous day

Next day
Watch us in action !

Our collections

Strains collection
Plasmids collection

Retrieved from "http://2012.igem.org/Team:Lyon-INSA/notebook"