Team:UNAM Genomics Mexico/prueba

From 2012.igem.org

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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 />
>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 />
<h2>09/15/12</h2><br />
>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 />
>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 ligated 97/98/99 psBIC3 [[Team:UNAM_Genomics_Mexico/Notebook/Protocols#Ligation | LIGATION PROTOCOL]]. <br />
>We ligated 97/98/99 psBIC3 [[Team:UNAM_Genomics_Mexico/Notebook/Protocols#Ligation | LIGATION PROTOCOL]]. <br />
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<img src='https://static.igem.org/mediawiki/2012/a/a8/UnamgenomicsoANDcadmio09_16_2012.jpg' height="300"/>
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<div class='captionaqua'>
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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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<img src='https://static.igem.org/mediawiki/2012/8/89/UnamgenomicsoANDcadmio09_16_2012_2.jpg' width="300"/>
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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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<img src='https://static.igem.org/mediawiki/2012/b/b0/UnamgenomicsoANDcadmio09_16_2012_3.jpg' height="300"/>
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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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</li>
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</ul>
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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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</li>
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</ul>
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<img src='https://static.igem.org/mediawiki/2012/3/38/UnamgenomicsoANDcadmio09_16_2012_5.jpg' height="300"/>
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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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<img src='https://static.igem.org/mediawiki/2012/d/d6/UnamgenomicsoANDcadmio09_18_2012.jpg' height="300"/>
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<div class='captionverde'>
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<p class='captionInside'>A3/Pveg lysis. <br /><br />
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<img src='https://static.igem.org/mediawiki/2012/8/8f/UnamgenomicsoANDcadmio09_18_2012_2.jpg' height="300"/>
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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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<img src='https://static.igem.org/mediawiki/2012/e/ec/UnamgenomicsoANDcadmio09_18_2012_3.jpg' height="300"/>
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<div class='captionverde'>
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<img src='https://static.igem.org/mediawiki/2012/f/fa/UnamgenomicsoANDcadmio09_18_2012_4.jpg' height="300"/>
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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>
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</li>
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</ul>
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<ul>
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<img src='https://static.igem.org/mediawiki/2012/9/94/UnamgenomicsoANDcadmio09_18_2012_5.jpg' height="300"/>
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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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</div>
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</li>
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</ul>
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</div>
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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