Abstract
The susceptibihty of RNA to degradation by exogenous and endogenous RNase activity following cell lysis has been well documented (1,2). Moreover RNA usually occurs complexed with protein from which it must be released. Precautions to be taken against exogenous RNase include the use of plastic gloves, autoclavmg solutions after adding 0.1% (v/v) diethyl pyrocarbonate (DEPC; except Trrs, which reacts), and baking glassware, spatulas, and so forth at 180°C overnight (3).
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Barnard E. A. (1964) The unfolding and refolding of rrbonuclease in urea solutions 1 Rates and extents of physical changes. J Mol Biol 10, 235–262
AVIV H. and Leder P. (1972) Purification of biologically active messenger RNA by chromatography on ollgothymidylic acidcellulose. Proc. Natl. Acad Sci USA 69, 1408–1412
Sambrook J., Fritsch E. F., and Mamatis T. (1989) Molecular Clonzng A Laboratory Manual 2nd ed., Cold Spring Harbor Laboratory Press, Cold Spring Habor, NY, pp. 7.3–7.5.
Blackbum P., Wilson G., and Moore S. (1977). Rrbonuclease inhibitor from human placenta. Purification and properties. J. Biol Chem. 252, 5904–5910.
Berger S L and Bnkenmerer C. S. (1979) Inhrbitron of intractable nucleases with ribonucleonde-vanadyl complexes. isolation of messenger ribonucleic acid from resting lymphocytes. Biochemistry 18, 5143–5149.
Jones P., Qiu J., and Rickwood D (1994) RNA isolation and analysis. BIOS Scientific Publishers, Oxford, UK, pp. 15–28.
Chirgwin J. M, Przybyla A. E., MacDonald R. J, and Rutter W. J. (1979) Isolatron of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry 18, 5294–5299.
MacDonell M. T., Hansen J. N., and Ortiz-Conde B. A. (1987) Isolation, purification and enzymatic sequencing of RNA. Methods in Microbiology 19, 357–404
Hilz H., Wiegers U., and Adamietz P. (1975) Stimulation of proteinase K action by denaturmg agents. Application to the isolation of nucleic acids and the degradation of masked proteins. Eur J. Biochem 56, 103–108
Lizardi P. M. (1983) Methods for the preparation of messenger RNA. Methods Enzymol 96, 24–38.
Chomczynski P. and Sacchi N. (1987) Single-step method of RNA isolation by acid guanidiniumthiocyanatephenolchloroform extraction. Anal. Biochem 162, 156–159.
Yamamoto K. R., Alberts B. M., Benzinger R., Lawhorne L., and Treiber G. (1970) Rapid bacteriophage sedimentation in the presence of polyethylene glycol and its application to large-scale virus purification. Virology 40, 734–744
Sambrook J., Fritsch E F, and Maniatis T. (1989) Molecular Cloning A Laboratory Manual 2nd ed., Cold Spring Harbor Laboratory Press, Cold Spring Habor, NY,pp. 1.23–1.41.
Jenné S., Miczka G., and Heptinstall J. (1993) Rapid extraction of bacterial ribosomal RNA with polyethylene glycol. Sixth European Congress on Biotechnology, Firenze, Italy, 3, WE022.
Albertsson P.-A (1971) Partition of Cell Particles and Macromolecules. 2nd ed., Wiley-Interscience, New York
Hengen P. N. (1996) Methods and Reagents. Trends Biochem Sci. 21, 112,113
Gillespie D. H., Cuddy K. K., Kolbe T., and Marks D. I. (1994) Dissolve and capture: a strategy for analysing mRNA in blood. Nature 367, 390,391.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1998 Humana Press Inc.
About this protocol
Cite this protocol
Heptinstall, J. (1998). Isolation of Total RNA from Bacteria. In: Rapley, R., Manning, D.L. (eds) RNA Isolation and Characterization Protocols. Methods in Molecular Biology™, vol 86. Humana Press. https://doi.org/10.1385/0-89603-494-1:47
Download citation
DOI: https://doi.org/10.1385/0-89603-494-1:47
Publisher Name: Humana Press
Print ISBN: 978-0-89603-494-5
Online ISBN: 978-1-59259-570-9
eBook Packages: Springer Protocols