The quantitative analysis of multiple mRNA species using oligonucleotide probes in an S1 nuclease protection assay.
The quantitative measurement of steady-state mRNA levels is fundamental to the analysis of gene expression. A variety of techniques are widely used to achieve this including Northern blotting, RNase protection, and S1 nuclease protection. We describe here in detail a relatively recent extension of the S1 nuclease protection technique (1) in which radiolabeled oligonucleotides are used as probes in a solution hybridization assay (2). The principle advantage of this technique is that it allows, in a single RNA sample, the simultaneous measurement of the relative levels of at least six mRNA species, including that of a control mRNA. Further, a large number of RNA samples can be analyzed at one time.
Tanay
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VA
Department of Pharmacology, Faculty of Medicine, University of Alberta, Edmonton, Canada.
Tancowny
B P
BP
Glencorse
T A
TA
Bateson
A N
AN
D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes; D012333:RNA, Messenger / Q000032:analysis*; D015719:Single-Strand Specific DNA and RNA Endonucleases / Q000378:metabolism*
D015345:Oligonucleotide Probes; D012333:RNA, Messenger; D015719:Single-Strand Specific DNA and RNA Endonucleases