RNA Carriers

Donald J Roufa droufa at matt.ksu.ksu.edu
Thu Apr 23 08:41:16 EST 1992


	Recently several people in my lab have experienced a common
problem in several unrelated RNase protection assays:  Specifically,
bacterial and/or yeast tRNA carrier added to pGEM P-32 transcripts
used as probes generate a large number of mainly small (<400 nt)
"background" protected bands with a wide assortment of probes
transcribed from several cloned genes.  Repurification of E coli tRNA
prior to use through guanidine isothiocyanate phenol and chloroform
extractions helps a bit particularly with respect to the larger
background bands.  However, even after exhaustive purification, a
complex array of smaller bands invariably are present, even in the
absence of cellular messenger RNAs (i.e. tRNA containing probe treated
with RNaseA + T1).  I seem to recall reading somewhere that others
frequently use glycogen as a carrier for their probes.  However, I
can't recall where I read that.  Can someone provide information about
the use of alternative macromolecular carriers, such as glycogen, for
radioactive RNA probes?  A reference to the literature or to a
commercial brochure would be greatly appreciated.

Don Roufa
-- 
    Don Roufa        DROUFA at MATT.KSU.KSU.EDU
Division of Biology  Kansas State Univ.   Manhattan, KS  66506           



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