Tricine buffer for PCR

Jim Owens jow at helix.nih.gov
Thu Dec 15 16:56:48 EST 1994


In article <3coqkb$ghu at usenet.ucs.indiana.edu> , ellenmq at ucs.indiana.edu
writes:
>50 mM			2-mercaptoethanol

Have you ever left out the mercaptoethanol?

It or another reducing agent are not used by PE-Cetus in any of their
buffer formulations.  It may not be necessary.

Traditionally 2-mercaptoethanol has been included in buffers for nick
translation because it was thought necessary for activity of DNA
polymerase I.  It is not.  There is only one cysteine in the monomer so a
disulfide bridge was not necessary for the molecule to be active.  But it
did not hurt either.

Good luck,

Jim Owens



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