Team:Northwestern/Notebook/Protocols/PCR Amplification

From 2011.igem.org

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We used [http://www.neb.com/nebecomm/products/productM0531.asp Phusion High Fidelity PCR Master Mix] from New England Biolabs
We used [http://www.neb.com/nebecomm/products/productM0531.asp Phusion High Fidelity PCR Master Mix] from New England Biolabs
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Assemble all reaction components on ice and quickly transfer the reactions to a thermocycler preheated to the denaturation temperature (98 degrees C). All components should be mixed and centrifuged prior to use. It is important to add Phusion Master Mix last in order to prevent any primer degredation.
Assemble all reaction components on ice and quickly transfer the reactions to a thermocycler preheated to the denaturation temperature (98 degrees C). All components should be mixed and centrifuged prior to use. It is important to add Phusion Master Mix last in order to prevent any primer degredation.
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For 50 uL reaction:
For 50 uL reaction:
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Revision as of 19:15, 23 August 2011

RETURN TO IGEM 2010


PCR Amplification


We used [http://www.neb.com/nebecomm/products/productM0531.asp Phusion High Fidelity PCR Master Mix] from New England Biolabs


Assemble all reaction components on ice and quickly transfer the reactions to a thermocycler preheated to the denaturation temperature (98 degrees C). All components should be mixed and centrifuged prior to use. It is important to add Phusion Master Mix last in order to prevent any primer degredation.


For 50 uL reaction:

10 uM forward primer 2.5 uL
10 uM reverse primer 2.5 μL
Template DNA variable
Nuclease-free water to final volume (including mix) of 50 μL
2X Phusion Master Mix 25 μL
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