PCR Restriction Site engineering
brunstei at unixg.ubc.ca
Mon Jan 24 15:06:16 EST 1994
In regards to potential problems with digestibility of the sites
one designs into the ends of PCR primers, I would reccomend checking out
the table "Cleavage close to the ends of DNA fragments" in the back of
NEB's catalog (p.182 in last year's edition, in case it's handy.) I have
relied on this when picking my sites and found it quite useful. I should
add the standard disclaimers that I am not paid by, associated with,
related to, or otherwise in contact with NEB....
As far as purification goes, I always just isolate the PCR product
from the reaction directly by agarose gel electrophoresis and
electroelution of the band, it's quick and works at lest for me.
Hope this helps.
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