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Team:PKU Beijing/Notebook/Protocol/PCR
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Revision as of 08:14, 7 October 2009 by
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Notebook
Notebook Home
Protocol
AND Gate 1
Input
Core
Bistable
AND Gate 2
Output
System Assembly
Notes By Person (PDF)
Protocol for PCR with EasyPfu DNA Polymerase
download PDF version
Preparation for Reaction Mixture
Reagent
Final concentration
Volume
10X Taq buffer
1X
2 µl
2.5 mM dNTP mix
0.25 mM
2 µl
Forward Primer
0.2-0.4 µM
1 µl
Reverse Primer
0.2-0.4 µM
1 µl
EasyPfu DNA Polymerase
25 units
1 µl
Template DNA
As required
<0.5 µg
ddH2O
to final volume
Cycling Conditions
Step
Temperature/℃
Time/min
Initial Denaturation
94
5
Denaturation
94
0.5
Primer Annealing
As required
As required
Extending
72
As required(0.5kb/min)
Final Extending
72
10
Number of Cycles
32-35
Cooling Down
4
10
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