View source for PCR Amplification with Platinum Taq High Fidelity

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[[ Category: Molecular Biology ]]
[[ Category: Cloning ]]
[[ Category: Genotyping ]]

==Materials==
* Platinum PCR SuperMix High Fidelity (Life Technologies cat# 12532-016)
* Amplification Primers (2 uM stock solution with both primers combined): 

==Amplification of Target Region==
* Using Platinum Taq High Fidelity amplify region of interest (design primers, should be <1000bp):
* Per reaction:
** 21.5 uL Platinum Taq High Fidelity Master Mix
** 2.5 uL of Primers from a 2 uM stock solution
** 1 uL of the Extracted DNA from the previous step.
* The PCR program should be
** 2 min at 95C
** 94C for 15
** 55C for 15s (adjust based on Tm of primers, use 5C less)
** 68C for 1min/kb amplicon
** Repeat steps 2-5 35X
** 68C for 10min