Kun:LabNotes/ExonomeSeq/2008-1-8

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Exp.1 Perform 2nd round PCR on the PAGE purified 450bp band from the previous capturing experiment.

                         dNTP       dUTP
    Template:            5ul       5ul
    10X PCR buffer:     10ul      10ul
    10mM dNTP:           2ul       2ul
    1mM dUTP:            0ul       1ul
    100uM AmpFSolV6.2  0.4ul     0.4ul
    100uM AmpRSolV6.2  0.4ul     0.4ul
    50X SYBG I:        0.4ul     0.4ul
    JumpStart Taq:       2ul       2ul
    H2O:                80ul      79ul

94C 3min -> (94C 30sec -> 72C 1min) x 10cyc -> 72C 3min -> 15C hold. Purify 50ul with Bioneer columns.

Exp.2 Perform another rounf of exon capture experiment

   Sheared NA10835(~300ng/ul)                3ul
   E55kv1 probes 107nM (01/07/08,purified)  17ul
   10X Ampligase buffer                   2.25ul

95C 10min -> 60C 40h -> add 2.5ul NSL mix -> 60C 1h -> (95C 2min -> 60C 1h) x 20 -> 37C 1min -> add 2ul exo I&III, 1ul T7 exo -> 37C 2h -> 95C 2min -> 4c hold