PCR with tricine buffer
Enrique Jose Labadan Frio - SCBC - 3421101
g3421101 at MUCC.MAHIDOL.AC.TH
Mon Feb 21 05:53:45 EST 1994
I PCRed a 3.5 kb gene using tricine instead of tris.Cl (tricine
is more resistant to pH changes with changes in temperature i.e., drastic
ones in PCR during different cycles of amplification). The buffer has
produced a clean band (no primer dimers nor other primer artefacts) and
is hypothesized to increase the processivity of the Taq DNA polymerase.
However, I'm having problems cloning this fragment. I've tried
Klenow fill-in and restricting the resulting fragment with RE's
introduced into the sequence via the primers.
HEEEEELLLPP! That's an understatement....
Thanks a lot
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