Team:UNICAMP-Brazil/Notebooks/October 4

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