General conditions for SSR markers
Primers are used at 0.035 microM (M13 tailed forward primer), 0.35 microM reverse primer and 0.175 microM M13 universal primer final concentration
dNTPs are used at 250 microM final concentration
MgCl2 concentration was generally 3.5 mM
In most cases 10 microliter reactions are suitable.
25-50 ng genomic DNA should be used per sample
General PCR conditions:
Denaturation at 96 degrees for 3 minutes followed by:
45 cycles of:
96 degrees 15 seconds
50 degrees 30 seconds
72 degrees 1 minute
followed by a final extension at 72 degrees for 7 minutes.
Samples are denatured in 50 microliter formamide stained with bromophenol blue and 0.5 microliter loaded on a Licor gel.
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