Kun:LabNotes/ASE/2007-12-6

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Revision as of 18:03, 6 December 2007 by KunZhang (talk | contribs) (New page: ==Exp.1 PCR on the pooled captured DNA== I still have some capturing reactions from 11-21-07, 11-26-07, 12-03-07 left. I decided to pool the reactions from the same templates togethers, an...)
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Exp.1 PCR on the pooled captured DNA

I still have some capturing reactions from 11-21-07, 11-26-07, 12-03-07 left. I decided to pool the reactions from the same templates togethers, and do PCR to make more Solexa sequencing library. The reason is that, based on the quantification experiment did yesterday, there are still not enough number of circularized molecules per target.

                              one reaction        8 reactions
          Template:           5~10ul                  each
          10x Buffer:          9.0ul                  72ul
          25mM MgCl2           5.4ul                43.2ul
          25mM dNTP            1.2ul                 9.6ul
          100uM AmpSolV6F2     0.4ul                 3.2ul
          100uM AmpSolV6R2     0.4ul                 3.2ul
          iTaq                   1ul                   8ul
          50X SYBG I           0.8ul                 6.4ul
          H2O                   72ul                 576ul

94C 3min -> 10 cycles of (94C 45sec -> 57C 1min -> 72C 1min) -> 12 cycles of (94C 45 sec -> 72 1min) -> 72C 3min -> 15C hold.