Ribonuclease Protection Assay Controls ?
leach at acs.bu.edu
Sat Dec 17 12:28:21 EST 1994
In article <3cu257$qgu at apakabar.cc.columbia.edu>,
jjm3 at sawasdee.cc.columbia.edu (Jeffrey Joseph Martino) wrote:
> OK here it goes: Doing Ribonuclease Protection Assays on total RNA
> from various mouse cells/tissues (Northern no signal,
> and initial RPAs - no signal)
> - actin controls give a good signal.
> Actin isn't too rare. Want to put a detection limit number on that
> RPA negative result.
> I thought adding known quantities of an RNA back to sample as a control
> would be OK to check detection limit, but I promised to keep an eye out
> for an endogenous alternative:
> Any ideas for a good (not tissue-specific) low level message (with
> probes available) for a control?
Adding labelled double stranded DNA is a good loading error/processing
GAPDH (metabolic enzyme) is available from clonetech...however, it does
vary with drug treatments etc...in our hands...we use b-actin/GAPDH
..... Martin Leach Email:leach at mbcrr.harvard.edu
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