*<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>
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
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.
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