pcr dna purification for sequencing
Chris LaRosa
clarosa at biocomp.unl.edu
Fri Aug 4 12:02:22 EST 2000
> Hello t
> Just my tuppence worth - i use Qiagen PCR purification spin columns (same
> kit as used to extract DNA from agarose gels).
>
> Can also use it for res. enzyme digest clean ups etc.
>
> Seems to work cleaning up pcr products for ABI based chemistry.
>
> Another method used in the past is Chromaspin columns from Clontech - bit
> more cumbersome and yeild can suffer. Also depends on the size of your pcr
> product.
>
The lab using ABI chemistries where I worked originally used qiagen spin
columns. They worked ok, but we found the gibco columns cheaper and
also that they yielded higher and more consistantly .
If you are going the do pcr sequencing , I think the protocols in a
previous post are very good. We spent months perfecting them for the
lab. (because of the high cost of the ABI regents , we spend alot of
time in the development fine tuning of the protocols.)
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