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