Kun:LabNotes/ExonomeSeq/2008-1-4
Jump to navigation
Jump to search
Colony PCR[edit]
Perform PCR on 48 colonies from E55kv1(250+450) for sequencing
2X Taq master mix: 15ul x 50 750 10uM M13 primer mix: 1ul 50 H2O: 14ul 700
94C 3min -> (94C 30sec -> 54C 30sec -> 72C 45sec) x 35 -> 72C 3min -> 4C hold.
I got 15 sequences of about the right size from the 2008-01-09 sequencing submission. 11/15 are on target, one is the dimer of a short insert, one is off-target capture by capturing arms of two different probes, one captured a SINE repeat on chr8, and the last one hit a wrong but highly similar target on the same chromosome.