RNA fingerprinting

Andrea D. Branch, Bonnie J. Benenfeld, Hugh D. Robertson

Research output: Contribution to journalArticlepeer-review

31 Scopus citations

Abstract

RNA fingerprinting analysis is the fastest way to obtain accurate information about an RNA processing reaction. The presence of a new spot, pGp, in one of the products identifies the exact phosphodiester bond cleaved during the processing reaction and indicates the final deposition of the phosphate group. The original RNA fingerprints were made using high-voltage electrophoresis on diethylaminoethyl (DEAE) paper for the second dimension. This fractionation procedure provides considerable information about the base composition of the oligonucleotides. The protocol chosen for ribonuclease digestion depends on whether the RNA is already radiolabeled. In all cases, RNAs are dried down in small silicon-treated glass tubes. Glass tubes are used because their clarity permits even minute quantities of RNA to be seen. The phosphatase treatment removes the 3′-phosphate groups produced by ribonuclease digestion. Samples are then sealed into drawn-out capillary tubes and are heated in a bath of boiling water for three minutes to inactivate the phosphatase.

Original languageEnglish
Pages (from-to)130-154
Number of pages25
JournalMethods in Enzymology
Volume180
Issue numberC
DOIs
StatePublished - 1 Jan 1989

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