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PMID: 1690560 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

A simple, efficient method for the separate isolation of RNA and DNA from the same cells.

BioTechniques ·Vol. 8 ·No. 2 ·1990-02-00 ·Pages 154-6

Nicolaides NC, Stoeckert CJ

Abstract

A method for the isolation of total cytoplasmic RNA and high molecular weight DNA from the same cells is described. Cells are gently lysed with NP40 in the presence of vanadyl ribonucleoside complex and the nuclei pelleted by centrifugation. RNA is purified by phenol/CHCl3 extraction of the lysate supernatant followed by ethanol precipitation. Protein is removed from high molecular weight DNA by salt-precipitation after nuclei are digested with proteinase K in the presence of sodium dodecyl sulfate. High yields of clean, intact RNA and DNA are obtained. A major advantage of the method is that it can be scaled down to quantitatively extract RNA and DNA from as little as 1000 cells.

MeSH Terms
Animals Cell Line Cytoplasm/analysis DNA/isolation & purification Fibroblasts Humans Methods Mice Molecular Weight RNA/isolation & purification Sodium Chloride Tumor Cells, Cultured
Chemicals
Sodium Chloride RNA DNA
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Nicolaides N C
Department of Anatomy, University of Pennsylvania, School of Medicine, Philadelphia 19104.
Stoeckert C J
Article Info
Journal
BioTechniques
Abbr.
Biotechniques
ISSN
0736-6205
Published
1990-02-00
Pages
154-6
Language
English
Region
England
NLM ID
8306785
Subset
IM
Analysis Services
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