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===Exp.1 eSNPs capture (cont.)=== Perform PCR on 5ul the capturing reaction on Hues6 gDNA, compare iTaq with JumpStart Taq. x 1 x 2 template: 5ul 10ul ABI 10x buffer: 4.5ul 9ul 25mM MgCl2: 3ul 6ul 10mM dNTP: 1ul 2ul 100uM AmpF2Sol: 0.15ul 0.3ul 100uM AmpRSol: 0.15ul 0.3ul 50X SYBG I: 0.4ul 0.8ul iTaq: 1ul 2x1ul H2O: 35.3ul 70.6ul 94C 3min -> 10 cycles of (94C 45sec -> 56C 2min -> 72C 1min) -> 10 cycles of (94C 45 sec -> 72C 1min) -> 72C 3min -> 15C hold. PAGE purification of ds amplicons, 3 lanes per sample, two 5-well 6% TBE gel in total. *Hues6 gDNA: 40ul of iTaq amplified and 40ul of JumpStart amplified DNA (did today); *Hues6 cDNA: 16ul of each of the five subsets from the first batch of amplicons using iTaq; *Hues6 cDNA(no mouse): 16ul of each of the five subsets from the first batch of amplicons using iTaq; PAGE quantification: [[Image:2001-11-12-eSNP84k-Hues6-seqlib.png|400px]] Hues6 gDNA: 14.5ng/ul = 468nM[WRONG] should be 181nM Hues6 cDNA: 11.5ng/ul = 371nM[WRONG] should be 144nM Hues6 no mouse cDNA: 15ng/ul = 484nM[WRONG] should be 187nM
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