Team:UNICAMP-Brazil/Notebooks/October 5

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(Difference between revisions)
(→finOP and Cre-Recombinase with pGEM strategy)
(→Colony PCR to confirm the BBa K112806 + BBa B0015 ligation)
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*<p style=”text-align:justify;”> A lot of colonies have grown, it looks like our ligation were ok</p>
*<p style=”text-align:justify;”> A lot of colonies have grown, it looks like our ligation were ok</p>
-
*<p style=”text-align:justify;”> We selected 10 colonies to a PCR colonies, the positive colonies will be used to do a innoculum.</p>
+
*<p style=”text-align:justify;”> We selected 10 colonies to a PCR colonies,we used BBa B0015 as our positive control.</p>
*<p style=”text-align:justify;”> All the colonies didn't have the insertion of BBa K112806, but the size of the fragment was ok to BBa B0015</p>
*<p style=”text-align:justify;”> All the colonies didn't have the insertion of BBa K112806, but the size of the fragment was ok to BBa B0015</p>
*<p style=”text-align:justify;”> We don't have material enough of BBa B0015 for other digestion, so we did another innoculum to do a miniprep tomorrow</p>
*<p style=”text-align:justify;”> We don't have material enough of BBa B0015 for other digestion, so we did another innoculum to do a miniprep tomorrow</p>

Revision as of 22:47, 20 October 2009

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ColiGuard

finOP and Cre-Recombinase with pGEM strategy

  • Today we performed minipreps from yesterday's inoculated cultures, according to Protocol 2.
  • We expect to recover pGEM plasmids finally containing singly our inserts for finO, finP and Cre-Recombinase w/o ATG.
  • As both sides of the inserts contains the A stick end, it's possible that we will find in some samples unwanted ligations (it could be ligate in an upside down position). Therefore, we will also need to perform PCRs to confirm that our inserts indeed are in the correct frame position.

Marcelo and Victor

Colony PCR to confirm the BBa K112806 + BBa B0015 ligation

  • A lot of colonies have grown, it looks like our ligation were ok

  • We selected 10 colonies to a PCR colonies,we used BBa B0015 as our positive control.

  • All the colonies didn't have the insertion of BBa K112806, but the size of the fragment was ok to BBa B0015

  • We don't have material enough of BBa B0015 for other digestion, so we did another innoculum to do a miniprep tomorrow

Marcos, Luige and Ane





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