Dumb question (?) re cycle sequencing of PCR products

Sasha Kraev bckraev at aeolus.ethz.ch
Wed Nov 9 10:36:10 EST 1994

It is just so happened that I have never heard of such a dogma, however, 
over time I seem to have come over an observation that it is much more
reliable to use primers with Tm>65 C if you intend to cycle sequence later.
It seems to relate to the suggestion of Guy Hoelzer, that DNA ends in a PCR
product may not be flush-ended. Hope this helps, Sasha

More information about the Methods mailing list