Since all of our fragments are correctly digested, we started today the ligation of both finO and finP sequences to the vector pSB1A3. Each sequence was ligated singly to the vector and, only after each one is confirmed, we'll join then both to a single construction.
We performed the ligation finO+vector pSB1A3 for an O/N period, according to Protocol 11.
Marcelo
finP+pSB1A3 ligation
As we did for finO sequence, we also ligated finP to the vector pSB1A3 according to the same protocol. The ligation also lasted O/N period.
Marcelo
PCR: Cre-Recombinase without ATG
Today we realized again the PCR reaction to isolate the Cre-Recombinase without the ATG codon. The amplification was succesfully realized, being comproved with an 1% agarose gel run. The photo proves the amplification and the expected size of our fragment:
YeastGuard
New biobricks in biobrick format
E. coli transformed with the new biobricks grew very well on the LB+AMP plates!
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