Estimation of total ribonucleic acid quantity from dilute samples by nondenaturing electrophoresis and silver staining

Often, the amount of RNA that can be isolated from a defined tissue is very small. A method consisting of nondenaturing polyacrylamide gel electrophoresis and silver staining is described that can be used to evaluate the concentration of very dilute RNA samples. The method is a good starting point f...

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Bibliographic Details
Published in:Electrophoresis 2000-02, Vol.21 (3), p.545-547
Main Authors: Kursula, Petri, Lehto, Veli-Pekka, Heape, Anthony M.
Format: Article
Language:English
Subjects:
Electrophoresis, Polyacrylamide Gel - methods
Humans
Nervous system
Nondenaturing electrophoresis
Protein Denaturation
RNA
RNA - analysis
Silver Staining
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Summary:Often, the amount of RNA that can be isolated from a defined tissue is very small. A method consisting of nondenaturing polyacrylamide gel electrophoresis and silver staining is described that can be used to evaluate the concentration of very dilute RNA samples. The method is a good starting point for assays dealing with small amounts of RNA, such as semiquantitative reverse transcription polymerase chain reaction (RT‐PCR), making it possible to perform parallel assays from similar amounts of total RNA when quantitation by other methods is too insensitive. The method has been used successfully to monitor the amount of total RNA isolated from rat sciatic nerve and the rat C6 glioma cell line.
ISSN:0173-0835
1522-2683
DOI:10.1002/(SICI)1522-2683(20000201)21:3<545::AID-ELPS545>3.0.CO;2-5