Beta-Agarase and PCR

r.nakisa at ic.ac.uk r.nakisa at ic.ac.uk
Fri Mar 5 16:22:22 EST 1993


Dear Netters,

I'm using beta-agarase to isolate a ligation product of two linear
fragments.  The fragments have sizes of 1000 and 400 b.p., and I cut
the 1400 ligation product out of a 1% LMP gel and agarase digest.
Then I spin down the undigested agarose, pipette the supernatent into
a new eppendorf and precipitate with isopropanol according to the
instructions.

But here's the crunch.  When I try to amplify the 1400 b.p. fragment,
it doesn't amplify at all!  Does anyone know whether Taq polymerase is
affected by the products of an agarase digestion?

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