Team:USTC CHINA/Notebook/Protocols/PCR

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Latest revision as of 08:57, 27 September 2013

PCR

Performed with TaKaRa PrimeSTAR® GXL DNA Polymerase

Volume (μl)
5×PrimeSTAR GXL Buffer 10
dNTP Mixture（2.5 mM each） 4
Forward primer 10~15 pmol
Reverse primer 10~15 pmol
Template DNA 10pg~10ng
PrimeSTAR GXL DNA Polymerase 1
ddH2O To 50
Total 50

3. Gently vortex the samples and spin down.
4. Perform PCR using recommended thermal cycling conditions:

Step Temperature, °C Time Number of cycles
Initial denaturation 98 8 min 1
Denaturation 98 10 s 25~40
Annealing 55(if Tm<60) 15 s 25~40
Annealing 60(if Tm>60) 15 s 25~40
Extension 68 1 min/kb 25~40
Final Extension 68 5-15 min 1

Notebook

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