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PMID: 2450333 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Rapid extraction of high molecular weight RNA from cultured cells and granulocytes for Northern analysis.

Nucleic acids research ·Vol. 16 ·No. 4 ·1988-02-25 ·Pages 1487-97

Birnboim HC

Abstract

The study of messenger RNA in mammalian cells by Northern analysis requires the extraction of intact RNA in pure form. Although a number of reliable techniques have been developed for the purpose, most are fairly complex, involving steps such as ultracentrifugation and multiple extractions with large volumes of phenol and chloroform. When the number of cell samples to be analyzed is large, these techniques can be unwieldy. I now describe an RNA purification procedure which is simple enough to allow handling of a large number of cultured cell samples. It uses safe and inexpensive reagents and produces a high yield of pure total cell RNA, essentially free of DNA and ribonuclease, suitable for Northern analysis. The procedure also allows extraction of intact RNA from human granulocytes, cells which are rich in ribonuclease and contain very low amounts of RNA.

MeSH Terms
Animals Cells, Cultured Electrophoresis, Agar Gel/methods Granulocytes/analysis Humans Indicators and Reagents Mice Nucleic Acid Hybridization RNA/blood,genetics,isolation & purification RNA, Messenger/genetics
Chemicals
Indicators and Reagents RNA, Messenger RNA
Authors & Affiliations
1 authors, click to expand affiliations / ORCID
Birnboim H C
Ottawa Regional Cancer Centre, University of Ottawa, Ontario, Canada.
References (12)
12 references, click to expand
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
Published
1988-02-25
Pages
1487-97
Language
English
Region
England
NLM ID
0411011
PMCID
PMC336329
Subset
IM
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