Denaturation of Streptavidin-Biotin bound dsDNA into ssDNA

Tom Knight via methods%40net.bio.net (by tk from shaggy.csail.mit.edu)
Mon Apr 12 12:09:53 EST 2010


Cathal Garvey <cathalgarvey from gmail.com> writes:
> As a possible improvement to Tom's approach, do a preceding PCR with a
> much larger product by using an upstream forward primer.

Easier is to biotinylate the opposite primer, do a normal pcr with
both primers, then a second reaction with only the forward primer.
This will leave the template with biotin, and the ssDNA desired
product with no label.  Pull out the biotinylated template, leaving
the desired ssDNA product pure.



More information about the Methods mailing list