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===Exp1: PCR debugging=== In this experiment, I'm setting up a number of PCR reactions with different template DNA, batches of primers, dNTPs and Taqs. The purpose is to find out what went wrong in the last three experiments. <br> <b> iTaq reactions</b> {| {{table}} | align="center" style="background:#f0f0f0;"|'''iTaq''' | align="center" style="background:#f0f0f0;"|'''1''' | align="center" style="background:#f0f0f0;"|'''2''' | align="center" style="background:#f0f0f0;"|'''3''' | align="center" style="background:#f0f0f0;"|'''4''' | align="center" style="background:#f0f0f0;"|'''5''' | align="center" style="background:#f0f0f0;"|'''6''' | align="center" style="background:#f0f0f0;"|'''7''' | align="center" style="background:#f0f0f0;"|'''8''' |- | Template, gDNA (10ng/ul)||HDAA50, 1ul||GM20431, 1ul||GM20431,cDNA, 1ul||HDAA50, 1ul||HDAA50, 1ul||GM20431, 1ul||GM20431,cDNA, 1ul||HDAA50, 1ul |- | 10X buffer||5ul||5ul||5ul||5ul||5ul||5ul||5ul||5ul |- | 50mM MgCl2||1.5ul||1.5ul||1.5ul||1.5ul||1.5ul||1.5ul||1.5ul||1.5ul |- | 10mM dNTP||ABI, 1ul||ABI, 1ul||ABI, 1ul||ABgene, 1ul||ABI, 1ul||ABI, 1ul||ABI, 1ul||ABgene, 1ul |- | 50X SYBG I||0.4ul||0.4ul||0.4ul||0.4ul||0.4ul||0.4ul||0.4ul||0.4ul |- | iTaq||0.25ul||0.25ul||0.25ul||0.25ul||0.25ul||0.25ul||0.25ul||0.25ul |- | H2O||40ul||40ul||40ul||40ul||40ul||40ul||40ul||40ul |- | 10uM Primer||rs1264899, 1ul||rs1264899, 1ul||rs1264899, 1ul||rs1264899, 1ul||rs1045216, 1ul||rs1045216, 1ul||rs1045216, 1ul||rs1045216, 1ul |- | |} <b>Thermo Start Taq</b> {| {{table}} | align="center" style="background:#f0f0f0;"|'''Thermo Start Taq''' | align="center" style="background:#f0f0f0;"|'''1''' | align="center" style="background:#f0f0f0;"|'''2''' | align="center" style="background:#f0f0f0;"|'''3''' | align="center" style="background:#f0f0f0;"|'''4''' | align="center" style="background:#f0f0f0;"|'''5''' | align="center" style="background:#f0f0f0;"|'''6''' | align="center" style="background:#f0f0f0;"|'''7''' | align="center" style="background:#f0f0f0;"|'''8''' |- | Template, gDNA (10ng/ul)||HDAA50, 1ul||GM20431, 1ul||GM20431,cDNA, 1ul||HDAA50, 1ul||HDAA50, 1ul||GM20431, 1ul||GM20431,cDNA, 1ul||HDAA50, 1ul |- | 10X buffer||5ul||5ul||5ul||5ul||5ul||5ul||5ul||5ul |- | 25mM MgCl2||3ul||3ul||3ul||3ul||3ul||3ul||3ul||3ul |- | 10mM dNTP||ABI, 1ul||ABI, 1ul||ABI, 1ul||ABgene, 1ul||ABI, 1ul||ABI, 1ul||ABI, 1ul||ABgene, 1ul |- | 50X SYBG I||0.4ul||0.4ul||0.4ul||0.4ul||0.4ul||0.4ul||0.4ul||0.4ul |- | Thermo Start Taq||0.25ul||0.25ul||0.25ul||0.25ul||0.25ul||0.25ul||0.25ul||0.25ul |- | H2O||38.5ul||38.5ul||38.5ul||38.5ul||38.5ul||38.5ul||38.5ul||38.5ul |- | 10uM Primer||rs1264899, 1ul||rs1264899, 1ul||rs1264899, 1ul||rs1264899, 1ul||rs1045216, 1ul||rs1045216, 1ul||rs1045216, 1ul||rs1045216, 1ul |- | |} <b>Thermocycling protocol:</b><br> 95C 15min (3min for iTaq reactions) -> 40 cycles of (94C 30S -> 56C 30S -> 72C 1min) -> 72C 3min <br>
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