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ColiGuard
finO and finP's biobricks confirmation
Today we performed a PCR of each September 15th's minipreps samples, using the finO and finP designed primers, in order to confirm that our inserts are correctly contained in the plasmid vector. Unfortunately, our lab ran out of Taq Polymerase, and we were forced to used a home-made Taq, which efficiency is much lower than the industrialized one.
Sadly, we weren't able to confirm finO and finP's biobricks, as our PCR's didn't amplified even a single fragment. However, that result may be a consequence of the home-made Taq usage, and we intend to repeat those PCR's with a trustable Taq.
Marcelo
Cre-Recombinase + pSB1A3 ligation
Once we digested Cre-Recombinase's insert and the vector with XbaI and SpeI restriction enzymes, we could perform the ligation between them, according to protocol Protocol 11.
Then, we transformed the ligation into electrocompetent 'E. coli' bacteria, strain DH10B, accordin to protocol Protocol 3.
After being incubated at 37ºC for 1 hour, transformed cells were plated into solid LB-AMP media, and were grown at 37ºC for an O/N period.
Víctor
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