Direct Sequencing of degenerated PCR products

[Tom Knight]

adrien <adrien.vetterli from helsinki.fi> writes:
> I would like to sequence PCR products obtained from a reaction with
> degenerated primers without having to go through the cloning
> thing. The primers to  be used in the sequencing reaction are
> degenerated as well. Has anyone tried that who can tell me if the
> results are usable ? I know of some scientists who have designed
> degenerated primers with SP6 or T7 extensions and use these sites
> during sequencing instead of the degenerated ones but I am not going
> there either.

I've done this with no special considerations.  Just gel isolation of
the PCR product band and addition of one of the degenerate primers.
You can't determine the specific bases of the degenerate primers you
have used by sequencing in the opposite direction -- they give mixed
or incorrect results for the degenerate positions.