RNA isolation and fractionation with compaction agents

Jason C. Murphy, George E. Fox, Richard C. Willson

Research output: Contribution to journalArticle

9 Scopus citations

Abstract

A new approach to the isolation of RNA from bacterial lysates employs selective precipitation by compaction agents, such as hexammine cobalt and spermidine. Using 3.5 mM hexammine cobalt, total RNA can be selectively precipitated from a cell lysate. At a concentration of 2 mM hexammine cobalt, rRNA can be fractionated from low molecular weight RNA. The resulting RNA mixture is readily resolved to pure 5S and mixed 16S/23S rRNA by nondenaturing anion-exchange chromatography. Using a second stage of precipitation at 8 mM hexammine cobalt, the low molecular weight RNA fraction can be isolated by precipitation. Compaction precipitation was also applied to the purification of an artificial stable RNA derived from Escherichia coli 5S rRNA and to the isolation of an Escherichia coli-expressed ribozyme.

Original languageEnglish (US)
Pages (from-to)143-148
Number of pages6
JournalAnalytical Biochemistry
Volume295
Issue number2
DOIs
StatePublished - Aug 15 2001

Keywords

  • Compaction
  • Purification
  • Ribozyme
  • RNA
  • rRNA
  • Selective precipitation
  • Separation

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Fingerprint Dive into the research topics of 'RNA isolation and fractionation with compaction agents'. Together they form a unique fingerprint.

Cite this