Team:UNICAMP-Brazil/Notebooks/October 4

From 2009.igem.org

(Difference between revisions)
(→Transformation of the BBa K112806 + BBa B0015 ligation)
 
(5 intermediate revisions not shown)
Line 5: Line 5:
==''' ColiGuard '''==
==''' ColiGuard '''==
 +
 +
====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>
 +
 +
''Victor''
====finOP and Cre-Recombinase with pGEM strategy====
====finOP and Cre-Recombinase with pGEM strategy====
Line 16: Line 23:
====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 eletroporation according to the [https://2009.igem.org/Team:UNICAMP-Brazil/Protocols/Electroporation  Protocol 3]</p>
+
*<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''
Line 27: Line 34:
''Fabi and Léo''
''Fabi and Léo''
-
 
==''' YeastGuard '''==
==''' YeastGuard '''==
-
====New strategy: pGEM====
+
====[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

Topo l2.gif topo_r_igem.gif
topo_r_b.gif
Back to Calendar

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




Retrieved from "http://2009.igem.org/Team:UNICAMP-Brazil/Notebooks/October_4"