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Detection and quantitation of specific mRNAs by ribonuclease protection assay using denaturing horizontal polyacrylamide gel electrophoresis: a radioactive and nonradioactive approach.

作者信息

Plath A, Peters F, Einspanier R

机构信息

Institut für Physiologie, Forschungszentrum für Milch und Lebensmittel, Weihenstephan, Freising, Germany.

出版信息

Electrophoresis. 1996 Mar;17(3):471-2. doi: 10.1002/elps.1150170306.

DOI:10.1002/elps.1150170306
PMID:8740160
Abstract

A radioactive (32P) and nonradioactive (digoxigenin) ribonuclease protection assay (RPA) has been developed to detect mRNAs of housekeeping proteins and growth factors. A modification of polyacrylamide gel electrophoresis (PAGE) to simplify RPA is described. Both Cleangels (Pharmacia) and laboratory-cast polyacrylamide gels, in a denaturing, horizontal electrophoresis system, were used. The amount of toxic chemicals and waste was reduced, in comparison with sequencing gels normally used for RPA. The protected RNA fragments were shown to be well-separated, with sufficient sensitivity in this modified, quick gel system.

摘要

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