Team:Peking S/lab/protocol/pcr

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<font size=6> <font color="#FFFFFF">Protocol</font></FONT>



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==='''Protocol for PCR with EasyPfu DNA Polymerase'''===

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'''Preparation for Reaction Mixture'''
{|cellpadding=5
|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'''
{|cellpadding=5
|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
|}