Direct Sequencing of RT-PCR products.

Helgi Briem Magnusson hbriem at rhi.hi.is
Tue Aug 24 06:20:50 EST 1993


In <1993Aug24.013048.29384 at gserv1.dl.ac.uk> N.SALAZAR at uk.ac.lshtm (Nelson Salazar) writes:


>Hi everybody,
>                I would also appreciate very much if someone could
>help us out with the sequencing of PCR products without having to
>clone them. I have used the Pharmacia kit, which seems quite simple,
>but I failed the first time. Now I am preparing more template DNA
>(ssDNA) and I hope that this time it will work. However, if someone
>has used this approach or has a better one, that would help us quite
>a bit.

I would like to recommend that you try Dynal's streptavidin coated
magnetic beads, Dynabeads (tm) together with a biotinylated primer.
It is not cheap, but it is extremely quick and easy and gives very
good results.  It is especially useful when the same region must
be sequenced over and over, as for population studies.  It is 
probably too expensive to use for a one-shot sequencing, as tagging
the primer with biotin is expensive and so are the beads themselves.

Helgi Briem
Inst. Exp. Path.
Keldur, Iceland



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