10 colonies from both ligations (pCR2.1/ORF20 & pCR2.1/ORF25) were picked with clean pipette tips, streaked on a different LB-Amp50+X-gal plate that had been gridded and numbered, and then used to inoculate the PCR reactions.
PCRs were performed with respective phage primers.
Master mix calcs are here: 20140909 – Phage Colony Screens
- 95C – 10mins
- 95C – 15s
- 55C – 15s
- 72C – 30s
Top half of gel are the pCR2.1/ORF20 colonies.
Bottom half of gel are the pCR2.1/ORF25 colonies.
All amplified. Will select one of each from the gridded plates for plasmid isolation for use in qPCR standard curves and in-situ hybridization (ISH).