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RNA isolation and fractionation with compaction agentsA 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. Copyright 2001 Academic Press.
Document ID
20040112341
Acquisition Source
Legacy CDMS
Document Type
Reprint (Version printed in journal)
External Source(s)
Authors
Murphy, J. C.
(University of Houston 4800 Calhoun Avenue, Houston, TX 77204-4792, United States)
Fox, G. E.
Willson, R. C.
Date Acquired
August 21, 2013
Publication Date
August 15, 2001
Publication Information
Publication: Analytical biochemistry
Volume: 295
Issue: 2
ISSN: 0003-2697
Subject Category
Life Sciences (General)
Report/Patent Number
ISSN: 0003-2697
Distribution Limits
Public
Copyright
Other
Keywords
Non-NASA Center
NASA Discipline Environmental Health
RNA, Bacterial/isolation & purification
Support, Non-U.S. Gov't
Escherichia coli
Chromatography, Ion Exchange
Cobalt
Precipitation
RNA, Ribosomal, 5S/isolation & purification
Support, U.S. Gov't, Non-P.H.S
Chemical Fractionation

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