Purifying 5' phosphorylated primers for mutagenesis

Haley Lindsey via methods%40net.bio.net (by haleyl from u.washington.edu)
Tue Nov 24 17:12:50 EST 2009


I'm doing some site-directed mutagenesis with a kit from NEB. It is
recommended in the kit directions that the 5' phosphorylated primers used
for the reaction are purified either using RP-HPLC or PAGE.

My issue is that these purification methods are *really* expensive. (I've
been getting my primers through Invitrogen, but I've also looked at a few
other places and it's the same story.) Has anyone had success using primers
purified another (less expensive) way or done the purification in house?

Thank you,

Haley, University of Washington


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