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==PCR== {| {{table}} | align="center" style="background:#f0f0f0;"|'''Content''' | align="center" style="background:#f0f0f0;"|'''Volume''' |- | PfuTurboCx Hotstart DNA Polymerase 2.5U/ul||1 |- | 10x PfuTurbo reaction buffer||5 |- | dNTPs 1mM each||10 |- | PCR F 10uM||1 |- | PCR R 10uM||1 |- | Template||2 |- | 50X SYBR Green||0.2 |- | H2o||29.8 |- | Total||50 |} 95 °C for 2 min, 98 °C for 30 s, then 29 cycles of 98 °C for 15 s, 60 °C for 30 s and 72 °C for 4 min, ending with one 72 °C for 10 min step [[File:methylone_120117.jpg]] *run gel to see the amplicon size [[File:ZhangLab_2 2012-01-30 16hr 54min.jpg|300px]] *1-after A-tailing *2-after adapter ligation *3-after bisulfite conversion and PCR *Dr. Zhang decides to switch the polymerase to iQ, it could be the SYBR Green that inhibits the amplification
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