Team:UNAM Genomics Mexico/prueba

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{{:Template:Team:UNAM_Genomics_Mexico/webhtml| content=
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= MAY =
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<html>
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<div class='thumbnailWrapper'>
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<ul>
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<img src='https://static.igem.org/mediawiki/2012/5/5c/UnamgenomicsoANDcadmio09_14_2012.jpg' width="300"/>
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<div class='captiongray'>
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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.CI 02 E/P. <br />
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5.P4 PCR. <br />
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</div>
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</li>
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</ul>
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<br /><br />
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<img src='https://static.igem.org/mediawiki/2012/0/04/UnamgenomicsoANDcadmio09_14_2012_2.jpg' height="300"/>
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<div class='captionnaranja'>
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<p class='captionInside'>We extracted the correct band by kit <br />
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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.CI 02 E/P. <br />
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5.P4 PCR. <br />
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</div>
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</li>
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</ul>
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<br /><br />
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<img src='https://static.igem.org/mediawiki/2012/e/ea/UnamgenomicsoANDcadmio09_14_2012_3.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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2.00179 Lasr lysis<br />.
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3.00179 LasR. <br />
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4.00179 Lasr (-).<br />
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5.LasR 00179 new primers. <br />
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6.LasR 00179 new primers. <br />
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7.LasR 00179 new primers (-).<br />
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<img src='https://static.igem.org/mediawiki/2012/9/9e/UnamgenomicsoANDcadmio09_14_2012_4.jpg' height="300"/>
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<div class='captionverde'>
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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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<img src='https://static.igem.org/mediawiki/2012/5/55/UnamgenomicsoANDcadmio09_14_2012_5.jpg' height="300"/>
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<div class='captionrosa'>
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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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</div>
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</li>
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</ul>
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</div>
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<br /><br />
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</html>
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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 />
 +
2.ArsR-CzRA 98 E/P. <br />
 +
3.1 kb ladder. <br />
 +
4.ArsR-CzRA 99 E/P. <br />
 +
5.CI 02 E/P. <br />
 +
6.AmyE 5’ 97 E/S. <br />
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7.AmyE 5’ 98 E/S. <br />
 +
8.1 kb ladder. <br />
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9.AmyE 5’ 99 E/S. <br />
 +
10.LasR PCR. <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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<br /><br />
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</html>
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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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<center><h1>'''Bacillus subtilis Results'''</h1></center>
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We know that everybody has wetlab problems, just like us. And we actually spend most of the summer looking to standardize the Bacillus subtilis transformation protocol. And we did it! So we are presenting you our new standard:
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[Team:UNAM_Genomics_Mexico/Notebook/Protocols#Escherichia_coli_MC1061_competent_cells_protocol Escherichia coli MC1061 competent cells protocol]
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[[Team:UNAM_Genomics_Mexico/Notebook/Protocols#Escherichia_coli_MC1061_competent_cells_protocol | Link title]]
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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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<table border="0"  height="150" cellspacing="15" bgcolor="transparent" cellpadding="10" id="tablecontentbg">
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<td  id="contentcolumn2" align="center"><iframe width="853" height="480" src="http://www.youtube.com/embed/rzj69X8n2ks" frameborder="0" allowfullscreen></iframe></td>
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<img src='https://static.igem.org/mediawiki/2012/a/a8/UnamgenomicsoANDcadmio09_16_2012.jpg' height="300"/>
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</table>
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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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</div>
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</li>
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</ul>
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<br /><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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</div>
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</li>
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</ul>
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</div>
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<br /><br />
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</html>
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<div class='thumbnailWrapper'>
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<ul>
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<li>
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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>
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</li>
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</ul>
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<br /><br />
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</html>
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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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</div>
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</li>
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</ul>
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</div>
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<br /><br />
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</html>
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<h2>09/16/12</h2><br />
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[[Team:UNAM_Genomics_Mexico/Notebook/Protocols#Escherichia_coli_MC1061_heat_shock_transformation_protocol Two-step Bacillus subtilis transformation procedure]]
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<td  id="contentcolumn2" align="center"><iframe width="853" height="480" src="http://youtu.be/6RxRGO8flqM" frameborder="0" allowfullscreen></iframe></td>
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We notice that you not only need a plasmid to transform into B. subtilis, but a plasmid that has form multimers in a RecA+ E. coli and should be flanked by AmyE 5' & 3', so that when the plasmid is transformed into Bacillus, it does not get degradated, and it makes an integration into the genome.
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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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<ul>
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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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<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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<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


Retrieved from "http://2012.igem.org/Team:UNAM_Genomics_Mexico/prueba"