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==Exp. 2 Perform CES22k eSNP capture on PGP and hybrid samples== {| border="1" cellpadding="5" cellspacing="0" align="center" |- | align="center" style="background:#f0f0f0;"|''' ''' | align="center" style="background:#f0f0f0;"|''' ''' | align="center" style="background:#f0f0f0;"|'''Template''' | align="center" style="background:#f0f0f0;"|'''10x buffer''' | align="center" style="background:#f0f0f0;"|'''CES22k probes (277nM)''' | align="center" style="background:#f0f0f0;"|'''H2O''' |- |1||GM20431 gDNA(347ng/ul)||2.3ul||1ul||5.4ul||0.3ul |- |2||GM20431 cDNA (121ng/ul)||0.4ul||1ul||5.4ul||2.2ul |- |3||GC1F cDNA(48ng/ul)||1ul||1ul||5.4ul||1.6ul |- |4||GC1EP cDNA(52ng/ul)||1ul||1ul||5.4ul||1.6ul |- |5||Hues6 gDNA(113ng/ul)||2.6ul||1ul||5.4ul||0ul |- |6||Hues6 cDNA(155ng/ul)||0.3ul||1ul||5.4ul||2.3ul |- |7||H1 gDNA(222ng/ul)||2.6ul||1ul||5.4ul||0ul |- |8||H1 cDNA(35ng/ul)||1.4ul||1ul||5.4ul||1.2ul |- |9||H2 gDNA(200ng/ul)||2.6ul||1ul||5.4ul||0ul |- |10||H2 cDNA(48ng/ul)||1ul||1ul||5.4ul||1.6ul |- |11||BJ MDA DNA(576ng/ul) ||2.6ul||1ul||5.4ul||0ul |- |12||BJ cDNA(18ng/ul)||2.6ul||1ul||5.4ul||0ul |} 94C 5min->60C 18h-> add 0.5ul SLN mix (1U/ul stoffel, 0.5U/ul AmpLigase, 100nM dNTP) -> 60C 4h -> 15 cycles of (94C 1min -> 60C 1h) -> 60C 4h -> 37C 1min -> add 1ul of exo mix (Exo I, Exo III, T7 exo)->37c 1 hour -> 94C 5min -> 4C hold. [11-22-2007] PCR one reaction 12 reactions Template: 5ul each 10x Buffer: 9.5ul 123.5ul 25mM MgCl2 6ul 78ul 25mM dNTP 1.2ul 15.6ul 100uM AmpSolV6F2 0.4ul 5.2ul 100uM AmpSolV6R2 0.4ul 5.2ul Platinium Taq 1ul 13ul 50X SYBG I 0.8ul 10.4ul H2O 76.7ul 997ul 94C 3min -> 10 cycles of (94C 45sec -> 57C 1min -> 72C 1min) -> 12 cycles of (94C 45 sec -> 72 1min) -> 72C 3min -> 15C hold. Added 3 more cycles for #3, 4, 6, 8, 10, 11, 12. [[Image:2007-11-24-CES22k-capture-PCR.png|500px]] Since the amplification curve for #1 came up much earlier than others, I repeat the PCR on 3ul #1 with 10 cycles at the second stage of amplification.
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