Cong T. Trinh:Taq PCR

Takara Taq polymerase/ Gene knockout detection

Reaction components

Chemical	Volume (μL)
LaTaq	        0.25
2xGC 1	        12.5
dNTP	        4
p1 (100 mM)	0.5
p2 (100 mM)	0.5
template*	2
water	        5.25
Total	        25

• 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) 	           94°C               5 min for colony PCR, 2 min for DNA	
Step 2.1     (30 cycle)    	   Td = 94°C                  30 sec	
Step 2.2     (30 cycle)	   Ta = Construct Dependent           30 sec	
Step 2.2     (30 cycle)           Text = 72°C                 2 min	
Step 3       (1 cycle)	           72°C                       7 min

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