Ran a colony PCR at the same time that I inoculated liquid cultures, using M13 primers.
Cycling params:
- 94C – 10m
40 cycles of:
- 94C – 1m
- 50C – 1m
- 72C – 2m
Results:
Lane 1: Hyperladder I
Lane 2: PGS Lo 1
Lane 3: PGS Hi 3
Lane 4: PGS Hi 4
Lane 5: Neg. Control
The only colony with an insert is PGS Hi 4. Will run a plasmid prep. However, this is the same sample that was sent for sequencing that produced nothing but vector sequence…