IGEM:Cambridge/2008/Notebook/Turing Pattern Formation/2008/08/15
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PCRing out promoters
- 4 different promoters : Pxyl, Pspc, Ppac, Pupp
- Make master mix enough for 4RXNs
Digest and Ligate Agr A and C to pSB4C5
Previous PCR products of Agr A and C were digested using Fermentas Fast Digest - EcoRI and PstI according to their protocol except a longer time was given to digest.
3uL of AgrA was used and 4.4 uL of AgrC was used and incubated for 40minutes.
10uL totaling to 1ug of pSB4C5 plasmid was also digested and incubated for 10 minutes.
The digestion was visualized on a 0.8% Agarose gel and produced expected sizes:
Plasmid: approx 3bk
Agr A: approx 700bp
Agr C: approx 1300bp