Sequencing PCR product by degenerateprimer

* * at alf1.ngate.uni-regensburg.de
Thu Aug 29 06:12:43 EST 1996


KHUANG at CHEMVX.TAMU.EDU wrote:

>Hi, Netters,
>Have you sequenced your PCR product by degenerate primer? Some people said
>if you increase the concentration of primer, it works. How about your experience?
>Thanks \
>Huang

Huang
I did a lot of sequencing of PCR products using the same degenerated
primers that I took for the initial amplification.
Sequencing was performed in a thermocycler with P33 labeled primers.
...worked great!
I think during the amplification process almost all kinds of
degenerated primers become incorporated into your product since one
mismatch doesn't matter while annealing. So they can accumulate in
every cycle.

Go ahead and try it
Mike





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