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Detection of messenger RNA and low-abundance heteronuclear RNA with single-stranded DNA probes produced by amplified primer extension labeling

PJ Brooks, MG Kaplitt, SP Kleopoulos, T Funabashi, CV Mobbs and DW Pfaff

Laboratory of Neurobiology and Behavior, Rockefeller University, New York, New York.

We describe a procedure for detection of low-abundance cellular RNAs by in situ hybridization histochemistry, using single-stranded DNA probes produced by amplified primer extension labeling with Taq polymerase. We have used this approach to detect a number of high- and low-abundance RNA species and have found it to be a simple and reproducible method of obtaining sensitive probes for in situ hybridization studies. For example, DNA probes generated by amplified primer extension labeling can detect low-abundance heteronuclear RNAs in individual neurons. Since this procedure does not involve recombinant DNA technology or microbiological facilities, it should prove useful to a wide variety of investigators studying the regulation of gene expression at the cellular level.

Volume 41, Issue 12, pp. 1761-1766, 12/01/1993
Copyright © 1993 by The Histochemical Society


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