Kun:LabNotes/ASE/2008-1-13
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Exp. 1 Perform CES22k eSNP capture on PGP samples[edit]
Genomic targets: 800ng/3pg per haploid genome = 2.67x10^5 copies = 2.22x10~-19 moles Probes: 2.22x10~-19 moles * 22,000 * 125 = 1.22x10^-12 moles.
Template | 10x buffer | CES22k probes #5 (246nM, 01/12/08) | H2O | ||
1 | GM20431 gDNA(347ng/ul) | 2.3ul | 1ul | 5ul | 1.7ul |
2 | GM20431 cDNA (122ng/ul) | 1.6ul | 1ul | 5ul | 2.4ul |
3 | GC1F cDNA(57ng/ul) | 3.5ul | 1ul | 5ul | 0.5ul |
4 | GC1EP cDNA(114ng/ul) | 1.8ul | 1ul | 5ul | 2.2ul |
94C 5min->60C 40h-> add 1.5ul SLN mix (1U/ul stoffel, 0.5U/ul AmpLigase, 200nM dNTP) -> 65C 1h -> 20 cycles of (94C 2min -> 65C 1h) -> 37C 1min -> add 1.0ul of exo mix (Exo I, Exo III)->37c 2 hour -> 95C 5min -> 4C hold.