Team:Chiba/protocol/PCR

>Protcol

PCR


 * Resuspend primer in Nuculease free water to 100 µM

100µL
 * PCR mix
 * 1) DNA template          1µL
 * 2) Fwd primer           10µL (final con. 10 pM)
 * 3) Rev primer           10µL
 * 4) 10x thermo pol buffer 10µL
 * 5) dNTP mix             10µL
 * 6) DNA pol.              1µL
 * 7) dH20                 58µL


 * PCR cycle

Start: 94 °C for 5 min. (melt) cycle:         melt:            1 min. anneal :          30 sec. cycle end: extension:72 °C for 3.5 min. 25 cycles 72 °C for 10 min store: keep at 6 °C forever