Looking for RT PCR protocol for bacteria
hyho at GATE.SINICA.EDU.TW
Fri Jul 18 00:21:21 EST 1997
Could Mr. Clark posts the protocols for all the netters. Thanks a lot.
On Wed, 16 Jul 1997, Dr. Duncan Clark wrote:
> In article <5qggta$dvb$2 at nntp.ucs.ubc.ca>, LAB <rewhlab at cbdn.ca> writes
> >Does anyone have a good RT PCR protocol for use with Gm. negative bacteria?
> >Someone told me that there is a single tube method floating around but I have
> >not been able to find it.
> Remember that the majority of bacterial mRNA's have extremely short
> half-lifes. I have a saved post from a couple of years ago that
> mentioned that by the time you have centrifuged down your cells their
> mRNA is greatly depleted. They found that mixing cells with 80%ethanol
> and DEPC sorted that out. I will email you a copy of their protocol for
> getting intact mRNA.
> The problem with being on the cutting edge is that you occasionally get
> sliced from time to time....
> Duncan Clark
> DNAmp Ltd.
> TEl/FAX 01252376288
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