Kun:LabNotes/ExonomeSeq/2008-1-4

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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.