Team:Calgary/Notebook/Protocols/Process4
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- | <p>We have a mastermix of | + | <p>We have a 2X mastermix of taq DNA polymerase, SYBR Green, and dNTPs. Primers will be stored at 100μM and used at final concentrations between 200-600nM. The PCR mix will be as follows:</p> |
<CENTER> | <CENTER> |
Revision as of 22:04, 27 September 2011
qRT-PCR
We have a 2X mastermix of taq DNA polymerase, SYBR Green, and dNTPs. Primers will be stored at 100μM and used at final concentrations between 200-600nM. The PCR mix will be as follows:
Component | Volume |
Quanta PCR master mix | 5 uL |
Primers (Fwd&Rev) at 600nM each | 3 uL |
cDNA | 2 uL |
Final Volume | 10 uL |
The following cylcing conditions are used.
40 cycles of: |
Melting curve analysis |