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PMID: 4866433 Published · ppublish English Journal Article

The preparation of ribosomal ribonucleic acid from whole bacteria.

The Biochemical journal ·Vol. 106 ·No. 4 ·1968-02-00 ·Pages 897-903

Robinson HK, Wade HE

Abstract

1. A simple method for the preparation of ribonuclease-free ribosomal RNA is described in which ribonuclease-deficient bacteria are treated with acetone and the RNA is extracted with phenol and purified by precipitating it with potassium acetate. The treatment with acetone appears to render the cell wall permeable to RNA but not to DNA during the extraction with phenol. The method thus avoids the need to disrupt the bacteria and greatly simplifies the subsequent purification. 2. The method has been used successfully with ribonuclease-deficient strains of Escherichia coli, Pseudomonas fluorescens and Staphylococcus epidermidis. The recovered purified RNA accounts for about 70% of the total ribosomal RNA and shows the normal sedimentation pattern of the 16s and 23s components in the analytical centrifuge.

MeSH Terms
Acetates Acetone DNA, Bacterial Escherichia coli Ethanol Hot Temperature Methods Phenols Pseudomonas RNA, Bacterial/analysis Ribosomes/analysis Staphylococcus Time Factors Ultracentrifugation
Chemicals
Acetates DNA, Bacterial Phenols RNA, Bacterial Acetone Ethanol
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Robinson H K
Wade H E
References (24)
24 references, click to expand
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Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
0264-6021
Published
1968-02-00
Pages
897-903
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1198593
Subset
IM
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