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Team:UNICAMP-Brazil/Notebooks/October 4
From 2009.igem.org
(→Transformation of the BBa K112806 + BBa B0015 ligation) |
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==''' ColiGuard '''== | ==''' ColiGuard '''== | ||
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| + | ====Cre-Recombinase without ATG==== | ||
| + | *<p style=”text-align:justify;”>The colonies grew up well! All of them were transparent.</p> | ||
| + | *<p style=”text-align:justify;”>We inoculated twenty colonies in liquid LB-AMP media in order to make a miniprep tomorrow.</p> | ||
| + | *<p style=”text-align:justify;">We let then grow for overnight at 37°C.</p> | ||
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| + | ''Victor'' | ||
====finOP and Cre-Recombinase with pGEM strategy==== | ====finOP and Cre-Recombinase with pGEM strategy==== | ||
| - | * Yesterday's transformed and plated cells grew in the media! We found white and blue colonies in the plate just as expected (due to the beta-galactosidase gene contained in pGEM vector). | + | *<p style=”text-align:justify;”>Yesterday's transformed and plated cells grew in the media! We found white and blue colonies in the plate just as expected (due to the beta-galactosidase gene contained in pGEM vector).</p> |
| - | * We selected 10 white colonies (those theorically contains our inserts in the correct position, since it interrupts the coding sequence of beta-galactosidase) from each plate and inoculated them into liquid LB-AMP media. | + | *<p style=”text-align:justify;”>We selected 10 white colonies (those theorically contains our inserts in the correct position, since it interrupts the coding sequence of beta-galactosidase) from each plate and inoculated them into liquid LB-AMP media.</p> |
| - | * Inocula were keep at 37ºC, under 250 rpm, for an O/N period. | + | *<p style=”text-align:justify;”>Inocula were keep at 37ºC, under 250 rpm, for an O/N period.</p> |
''Fabi, Leo, Marcelo and Victor'' | ''Fabi, Leo, Marcelo and Victor'' | ||
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====Transformation of the BBa K112806 + BBa B0015 ligation==== | ====Transformation of the BBa K112806 + BBa B0015 ligation==== | ||
| - | *<p style=”text-align:justify;”>We transformed the ligation made yesterday by | + | *<p style=”text-align:justify;”>We transformed the ligation made yesterday by electroporation according to the [https://2009.igem.org/Team:UNICAMP-Brazil/Protocols/Electroporation Protocol 3]</p> |
''Léo'' | ''Léo'' | ||
| + | ==== PY Promoter - Transformation ==== | ||
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| + | *<p style=”text-align:justify;”>Today we transformed the ligations PY1 + pGEM and PY2 + pGEM into electrocompetent ''E. coli'' bacteria, strain DH10B. We followed [https://2009.igem.org/Team:UNICAMP-Brazil/Protocols/Electroporation Protocol 3] (Electroporation) without modifications.</p> | ||
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| + | *<p style=”text-align:justify;”>After the transformation we plated the transformed cells in LB-AMP-Xgal plates, and let them grow at 37ºC for an O/N period.</p> | ||
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| + | ''Fabi and Léo'' | ||
==''' YeastGuard '''== | ==''' YeastGuard '''== | ||
| - | ====New | + | ====[https://2009.igem.org/Team:UNICAMP-Brazil/Protocols/pGEMStrategy New Strategy: pGEM]==== |
| - | *<p style=”text-align:justify;”> | + | *<p style=”text-align:justify;”>Only the plates containing ''E. coli'' transformed with pDLD and Lysozyme showed colonies. Tomorrow we will make colony PCR to collect the right ones.</p> |
Latest revision as of 03:46, 22 October 2009
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