Team:UNAM Genomics Mexico/prueba

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<p class='captionInside'>1.ArsR-CzrA 97 E/P. <br />
<p class='captionInside'>1.ArsR-CzrA 97 E/P. <br />
2.ArsR-CzrA 98 E/P. <br />
2.ArsR-CzrA 98 E/P. <br />
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4.CI 02 E/P. <br />
4.CI 02 E/P. <br />
5.P4 PCR. <br />
5.P4 PCR. <br />
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4.CI 02 E/P. <br />
4.CI 02 E/P. <br />
5.P4 PCR. <br />
5.P4 PCR. <br />
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<p class='captionInside'>1. 1 kb ladder. <br />
<p class='captionInside'>1. 1 kb ladder. <br />
2.00179 Lasr lysis<br />.
2.00179 Lasr lysis<br />.
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6.LasR 00179 new primers. <br />
6.LasR 00179 new primers. <br />
7.LasR 00179 new primers (-).<br />
7.LasR 00179 new primers (-).<br />
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<p class='captionInside'>1.1 kb ladder. <br />
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3.ArsR-CzrA 97 E/P. <br />
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5.ArsR-CzrA 98 E/P. <br />
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7.ArsR-CzrA 99 E/P. <br />
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9.CI 02 E/P. <br />
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11.AmyE 5’ ArsR-CzrA 97 E/S. <br />
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13.AmyE 5’ ArsR-CzrA 98 E/S. <br />
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15.AmyE 5’ ArsR-CzrA 99 E/S. <br />
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<p class='captionInside'>1.ArsR-CzrA 97 E/P. <br />
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2.ArsR-CzRA 98 E/P. <br />
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3.1 kb ladder. <br />
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4.ArsR-CzRA 99 E/P. <br />
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5.CI 02 E/P. <br />
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6.AmyE 5’ 97 E/S. <br />
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7.AmyE 5’ 98 E/S. <br />
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8.1 kb ladder. <br />
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9.AmyE 5’ 99 E/S. <br />
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10.LasR PCR. <br />
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<img src='https://static.igem.org/mediawiki/2012/2/2f/UnamgenomicsoANDcadmio09_14_2012_6.jpg' height="300"/>
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<div class='captionaqua'>
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<p class='captionInside'>1.ArsR-CzrA 97 E/P. <br />
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2.ArsR-CzRA 98 E/P. <br />
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3.1 kb ladder. <br />
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4.ArsR-CzRA 99 E/P. <br />
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5.CI 02 E/P. <br />
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6.AmyE 5’ 97 E/S. <br />
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7.AmyE 5’ 98 E/S. <br />
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8.1 kb ladder. <br />
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9.AmyE 5’ 99 E/S. <br />
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10.LasR PCR. <br />
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<h2>09/14/12</h2><br />
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We extracted the correct band by kit [[Team:UNAM_Genomics_Mexico/Notebook/Protocols#Gel_extraction_protocol | GEL EXTRACTION PROTOCOL]]. <br /><br />
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>We transformed ligation psB1c3+ArsR-CzrA 97/98/99/CI in competent DH5α cells from the pBad/pXyl AND team and in our own DH5α competent cells[[Team:UNAM_Genomics_Mexico/Notebook/Protocols#Transformation | TRANSFORMATION PROTOCOL]]. <br /><br />
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<h2>09/15/12</h2><br />
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>We checked our transformations an did liquid cultures and passed the colonies to new plates [[Team:UNAM_Genomics_Mexico/Notebook/Protocols#Liquid_culture | LIQUID CULTURE PROTOCOL]]. <br />
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>We digested E0040+B0014 [[Team:UNAM_Genomics_Mexico/Notebook/Protocols#Digestion_protocol_.2820_.C2.B5l.29 | DIGESTIONS PROTOCOL]]. <br />
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>We ligated 97/98/99 psBIC3 [[Team:UNAM_Genomics_Mexico/Notebook/Protocols#Ligation | LIGATION PROTOCOL]]. <br />
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<p class='captionInside'>1.AmyE 5’ 97 S/P. <br />
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2.
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3.AmyE 5’ 98 S/P. <br />
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4.
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5.1 kb ladder. <br />
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6.AmyE 5’ 99 S/P. <br />
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7.AmyE 5’ 99 S/P. <br />
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8.E0040 B0014 X/P. <br />
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9.
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<div class='captiongray'>
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<p class='captionInside'>1.E0040 B0014 X/P to extract. <br />
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2.1 kb ladder. <br />
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3.Lysis. <br />
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4.-19. Amy 5’ 99/97/P4CIE1010B0014 lysis. <br />
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<div class='captionnaranja'>
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<p class='captionInside'>1.LasR PCR. <br />
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2.LasR PCR. <br />
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3.LasR PCR. <br />
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<img src='https://static.igem.org/mediawiki/2012/c/c1/UnamgenomicsoANDcadmio09_16_2012_4.jpg' height="300"/>
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<div class='captionrojo'>
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<p class='captionInside'>1.1 kb ladder. <br />
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2.Purified P4. <br />
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3.Purified LasR. <br />
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<div class='captionverde'>
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<p class='captionInside'>Top part is degraded.
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Bottom:
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1.99 E/P. <br />
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2.99 E/P. <br />
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3.CI 02 E/P. <br />
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4.CI 02 E/P. <br />
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<h2>09/16/12</h2><br />
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<h2>05/29/2012</h2><br />
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Our group is in charge of building part of the “and” construction. We started analyzing if the plasmid we have with P4 actually had what we needed. <br /><br />
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The plasmid PRMn25 contains the protein P4. It has Amp100  resistance and comes in ''Escherichia coli''  NFI. Cells were lysed to make sure the plasmid was present in these cells (5000bp). We ran a gel with 3 lysis, a sample from the plasmid PFRC54 (A3 promoter) and a sample of total DNA from the strain from which PFRC54 was obtained. <br /><br />
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<p class='captionInside'>A3/Pveg lysis. <br /><br />
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<div class='captionverde'>
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<p class='captionInside'>1.-12. psCIC3 omega lysis. <br />
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13. 1 kb ladder. <br />
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14. psBIc3-GusA. <br />
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<div class='captionverde'>
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<p class='captionInside'>Digested A3 and pVeg with E/P. <br />
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<div class='captionverde'>
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<p class='captionInside'>1.ArsR-CzrA 97 E/P. <br />
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2.ArsR-CzrA 98 E/P. <br />
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3.ArsR-CzrA 99 E/P. <br />
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4.02-CI E/P. <br />
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5.PCR P4 purified. <br />
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<h2>09/18/12</h2><br />
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Latest revision as of 06:47, 26 October 2012


UNAM-Genomics_Mexico

= MAY =

  • 1.ArsR-CzrA 97 E/P.
    2.ArsR-CzrA 98 E/P.
    3.ArsR-CzrA 99 E/P.
    4.CI 02 E/P.
    5.P4 PCR.



  • We extracted the correct band by kit
    1.ArsR-CzrA 97 E/P.
    2.ArsR-CzrA 98 E/P.
    3.ArsR-CzrA 99 E/P.
    4.CI 02 E/P.
    5.P4 PCR.



  • 1. 1 kb ladder.
    2.00179 Lasr lysis
    . 3.00179 LasR.
    4.00179 Lasr (-).
    5.LasR 00179 new primers.
    6.LasR 00179 new primers.
    7.LasR 00179 new primers (-).



  • 1.1 kb ladder.
    3.ArsR-CzrA 97 E/P.
    5.ArsR-CzrA 98 E/P.
    7.ArsR-CzrA 99 E/P.
    9.CI 02 E/P.
    11.AmyE 5’ ArsR-CzrA 97 E/S.
    13.AmyE 5’ ArsR-CzrA 98 E/S.
    15.AmyE 5’ ArsR-CzrA 99 E/S.



  • 1.ArsR-CzrA 97 E/P.
    2.ArsR-CzRA 98 E/P.
    3.1 kb ladder.
    4.ArsR-CzRA 99 E/P.
    5.CI 02 E/P.
    6.AmyE 5’ 97 E/S.
    7.AmyE 5’ 98 E/S.
    8.1 kb ladder.
    9.AmyE 5’ 99 E/S.
    10.LasR PCR.



  • 1.ArsR-CzrA 97 E/P.
    2.ArsR-CzRA 98 E/P.
    3.1 kb ladder.
    4.ArsR-CzRA 99 E/P.
    5.CI 02 E/P.
    6.AmyE 5’ 97 E/S.
    7.AmyE 5’ 98 E/S.
    8.1 kb ladder.
    9.AmyE 5’ 99 E/S.
    10.LasR PCR.



Contents

09/14/12


We extracted the correct band by kit GEL EXTRACTION PROTOCOL.

>We transformed ligation psB1c3+ArsR-CzrA 97/98/99/CI in competent DH5α cells from the pBad/pXyl AND team and in our own DH5α competent cells TRANSFORMATION PROTOCOL.















09/15/12


>We checked our transformations an did liquid cultures and passed the colonies to new plates LIQUID CULTURE PROTOCOL.
>We digested E0040+B0014 DIGESTIONS PROTOCOL.

>We ligated 97/98/99 psBIC3 LIGATION PROTOCOL.

  • 1.AmyE 5’ 97 S/P.
    2. 3.AmyE 5’ 98 S/P.
    4. 5.1 kb ladder.
    6.AmyE 5’ 99 S/P.
    7.AmyE 5’ 99 S/P.
    8.E0040 B0014 X/P.
    9.



  • 1.E0040 B0014 X/P to extract.
    2.1 kb ladder.
    3.Lysis.
    4.-19. Amy 5’ 99/97/P4CIE1010B0014 lysis.



  • 1.LasR PCR.
    2.LasR PCR.
    3.LasR PCR.



  • 1.1 kb ladder.
    2.Purified P4.
    3.Purified LasR.



  • Top part is degraded. Bottom: 1.99 E/P.
    2.99 E/P.
    3.CI 02 E/P.
    4.CI 02 E/P.



09/16/12



















  • A3/Pveg lysis.



  • 1.-12. psCIC3 omega lysis.
    13. 1 kb ladder.
    14. psBIc3-GusA.





  • Digested A3 and pVeg with E/P.




  • 1.ArsR-CzrA 97 E/P.
    2.ArsR-CzrA 98 E/P.
    3.ArsR-CzrA 99 E/P.
    4.02-CI E/P.
    5.PCR P4 purified.



09/18/12


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