probe labeling

Lab path02 at ftn.com
Wed Jan 21 20:33:33 EST 1998



Michael Flores wrote:

> I have been having major problems trying to label a particular probe.  I
> first attempted to label a PCR amplifed fragment that I gel purifed.
> This fragment did not label at all.  I confirmed there was DNA present
> by both spectrophotometric determination and electrophoresis of an
> aliquot of the purified fragment.  I next decided to digest the insert
> out of the plasmid.  I then gel purifed this and attempted to label the
> fragment.  The incorporation was very poor.  I have used 2 different
> kits (Gibco RadPrime, and Stratagene Prime-It2) for each method with no
> luck.  I would appreciate any suggestions.
>
> Mike Flores
> mf43046 at glaxowellcome.com

Hey Mike,
Try Pharmacia's Oligolabeling kit. Seems to work well for us.  Do you think
there's a problem with your column that you're using to purify the probe
with?

Sue





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