hot start in PCR(cycle seq)

Sasha Kraev bckraev at aeolus.ethz.ch
Sun Oct 30 06:04:28 EST 1994


I used fmol kit and a couple of other similar kits right as the protocols
say, moreover, the completed reaction can be left at r.t. overnight ( my cycler
has no refrigeration, and I think it is unnecessary, unless one uses one of 
these new polymerases with 3'-exo-activity), however, making cycle sequencing 
with degenerate primers, if I understood the very first question correctly, 
is a matter of pure luck, and the chances for success are small, since in a 
single sided PCR the amplification is linear and the reaction specificity 
drops considerably, compared to normal PCR.



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