Today we prepared electrocompetent E. coli to use in our transformations according to Protocol 4.
ColiGuards
Cre-Recombinase without ATG
Today we started the construction of Cre-Recombinase without ATG, in order to made a new biobrick involving this construction. We ressuspended the biobrick of Cre-Recombinase: BBa_J61047 and transformed in E. coli DH10B eletrocompetent cells. We then plated on LB Amp plate, according to information from registry page.
Víctor
PY Promoter - PCR amplification
Today we performed the PCR amplification of the PY promoter from the F plasmid previously extracted from the conjugative strain (July 16). We did two different reactions, using two types of forward primers (Ppy-F-1 and Ppy-F-2) for the same reverse primer (Ppy-R). The product amplified with the Ppy-F-1/Ppy-R primers has 133 bp and the one amplified with the Ppy-F-2/Ppy-R has 75 bp. We called these products PY1 and PY2 respectively. The sizes of the products were confirmed by an agarose gel:
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