Matt Gethers/CRI, Thailand/Labwork/PCRs/PCR Pfu Template

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PCR PFU Template

Rxn Mix

Reagent Volume/Amount
Paeru Genomic DNA Template 0.5 μl from So Pa Pan stock
Pfu 0.5 μl
4 μl 1 μM Stock
4 μl 1 μM Stock
dNTPs 1 μl 10 mM Stock
DMSO 2.5 μl
Buffer S 5 μl
dH20 32.5 μl
Total 50 μl


Rxn Conditions

Annealing Temperature: oC

Extension Time:

Cycle

Step Temperature Duration Notes
Initial Denaturation 95oC 3 minutes
Repeat Cyclic Steps 35x
Cyclic Denaturation 94oC 40 Seconds
Cyclic Annealing oC 40 Seconds
Cyclic Extension 72oC minutes
Repeat Cyclic Steps 35x
Final Extension 72oC 10 minutes

Run Notes

References