Cong T. Trinh:Pfu ultra hotstart

Pfu Ultra hotstart gene amplification

Reaction components

Chemical	Volume (μL)
Master Mix	25
p1 (100mM)	0.5
p2 (100 mM)	0.5
template     (10-15 ng/ul) 2
water	        22
Total         50

• If the template is plasmid, the concentration should be 10-15 ng/uL

Reaction cycles

PCR reaction settings

Step #       # of cycles    Temperature                     Time
Step 1       (1 cycle) 	        95°C               2 min	
Step 2.1     (30 cycle)    	Td = 95°C                  20 sec	
Step 2.2     (30 cycle)	Ta = Construct Dependent           20 sec	
Step 2.2     (30 cycle)        Text = 72°C                 30 sec/kb	
Step 3       (1 cycle)	        72°C                       5 min

---

Return To Protocols