General protocol for PCR
Each person uses a slightly different ratio of reagents.
For DNA amplicons up to 500 bp, use 25 uL reaction volumes as follows:
On ice add to thin-walled pcr tubes
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0.125 uL Taq polymerase (1.25 U, Gibco)
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2.5 uL of 10 x PCR buffer minus Mg (1x)
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0.75 uL of 50 mM MgCl2 (1.5 mM)
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x uL template DNA
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0.5 uL of 10 mM dNTP mix (0.2 mM)
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1.25 uL 10 uM primer mix (0.5 uM)
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water to 25 uL
If DNA amplicon is >500 bp, scale-up the reactions for 50 or 100 uL total
volume. Program thermocycler appropriately and perform PCR reactions.