RT-PCR

[Jeff Fairman]

Most RT reactions will work in a standard PCR buffer, but not all.  We use a
custom master mix with Taqman buffer A (see Sept. BioTechniques for details)
and have no problem using MuMLV in a Taqman one mixture qRT-PCR.  This same
reaction seems to have trouble with the PE Universal master mix.

You might try using a Taq gold or the antibody to inactivate Taq at low
temperatures if you are getting extra bands.


Jeff Fairman

-----------------------------------------------------------------------
Jeff Fairman, Ph.D.
Senior Scientist, Pharmacogenomics Research
Clingenix, Inc.
871 Industrial Road, Suite J
San Carlos, CA 94070
(650) 598-7645 (office)
(650) 598-7641 (fax)
jfairman at clingenix.com


visit:  http://thelabrat.netfirms.com - By Scientists... For Scientists.

Steven Kirov wrote in message <38393346.78133C5B at ns.medfac.acad.bg>...
>Can anybody tells me if I can perform effective reverse trancriptase
>reaction in PCR buffer (Boehringer for example) with AMV or MuMLV and
>how? I have some experience in this but I don't get very good resuls
>