Team:UNICAMP-Brazil/Notebooks/October 4

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====Cre-Recombinase without ATG====
====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;”>The colonies grew up well! All of them were transparent.</p>
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*<p style=”text-align:justify;”>We inoculated twenty colonies in liquid LB-Amp media in order to make a miniprep tomorrow.</p>
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*<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>
*<p style=”text-align:justify;">We let then grow for overnight at 37°C.</p>
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====Transformation of the BBa K112806 + BBa B0015 ligation====
====Transformation of the BBa K112806 + BBa B0015 ligation====
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*<p style=”text-align:justify;”>We transformed the ligation made yesterday by eletroporation according to the [https://2009.igem.org/Team:UNICAMP-Brazil/Protocols/Electroporation  Protocol 3]</p>
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*<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''
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==''' YeastGuard '''==
==''' YeastGuard '''==
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====New strategy: pGEM====
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====[https://2009.igem.org/Team:UNICAMP-Brazil/Protocols/pGEMStrategy New Strategy: pGEM]====
*<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>
*<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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ColiGuard

Cre-Recombinase without ATG

  • The colonies grew up well! All of them were transparent.

  • We inoculated twenty colonies in liquid LB-AMP media in order to make a miniprep tomorrow.

  • We let then grow for overnight at 37°C.

Victor

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).

  • 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.

  • Inocula were keep at 37ºC, under 250 rpm, for an O/N period.

Fabi, Leo, Marcelo and Victor

Transformation of the BBa K112806 + BBa B0015 ligation

  • We transformed the ligation made yesterday by electroporation according to the Protocol 3

Léo

PY Promoter - Transformation

  • Today we transformed the ligations PY1 + pGEM and PY2 + pGEM into electrocompetent E. coli bacteria, strain DH10B. We followed Protocol 3 (Electroporation) without modifications.

  • After the transformation we plated the transformed cells in LB-AMP-Xgal plates, and let them grow at 37ºC for an O/N period.

Fabi and Léo

YeastGuard

New Strategy: pGEM

  • Only the plates containing E. coli transformed with pDLD and Lysozyme showed colonies. Tomorrow we will make colony PCR to collect the right ones.


Raíssa and Taís




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