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Team:Northwestern/Notebook/Protocols/PCR Amplification
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|align = "center" |Nuclease-free water | |align = "center" |Nuclease-free water | ||
| - | |align = "center" | | + | |align = "center" |to final volume (including mix) of 50 μL |
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|align = "center" |2X Phusion Master Mix | |align = "center" |2X Phusion Master Mix | ||
Revision as of 19:14, 23 August 2011
PROJECT
RESULTS
CONSIDERATIONS
ABOUT US
NOTEBOOK
ATTRIBUTIONS
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
