Team:UNICAMP-Brazil/Notebooks/September 24

ColiGuard

E. coli transformation with F plasmid

• E. coli DH10β electrocompetent cells were transformed using the recircularized F plasmid, according to Protocol 3.

• After the incubation time, 100 μl of medium containing the transformed cells were plated in LB-AMP-Xgal culture plates, and incubated O/N at 37°C.

Gabriel

finO and finP - Still Trying to Confirm our Biobricks

• After the miniprep procedure, we performed PCRs with finO and finP's designed primer, in order to confirm that our inserted fragment is indeed contained in the vector.

• As we had already got an amplified fragment of expected size from those colonies, we expect that those PCRs results in a successful amplification of our fragments. Moreover, as we aimed in reducing the amount of genomic DNA, we should got less inespecific amplifications.

Marcelo

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

CeaB and CeiB: Miniprep and transformation

• Today, we made Miniprep of the transformated cells and we made PCR again, so that avoids unspecific amplification. We didn’t get the band size expected again.

Luige