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A Method for Extracting RNA from Dormant and Germinating Bacillus subtilis Strain 168 Endospores

Moeller, Ralf and Horneck, Gerda and Rettberg, Petra and Mollenkopf, H.-J. and Stackebrandt, E. and Nicholson, W.L. (2006) A Method for Extracting RNA from Dormant and Germinating Bacillus subtilis Strain 168 Endospores. CURRENT MICROBIOLOGY, 53, pp. 227-231. Springer Science+Business Media.

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Abstract

RNA was extracted from dormant and germinating Bacillus subtilis 168 spores (intact spores and chemically decoated spores) by using rapid rupture followed by acid–phenol extraction. Spore germination progress was monitored by assaying colony forming ability before and after heat shock and by reading the optical density at 600 nm. The purity, yield, and composition of the extracted RNA were determined spectrophotometrically from the ratio of absorption at 260 nm to that at 280 nm; in a 2100 BioAnalyzer, giving the RNA yield/108 spores or cells and the distribution pattern of rRNA components. The method reported here for the extraction of RNA from dormant spores, as well as during different phases of germination and outgrowth, has proven to be fast, efficient and simple to handle. RNA of a high purity was obtained from dormant spores and during all phases of germination and growth. There was a significant increase in RNA yield during the transition from dormant spores to germination and subsequent outgrowth. Chemically decoated spores were retarded in germination and outgrowth compared with intact spores, and less RNA was extracted; however, the differences were not significant. This method for RNA isolation of dormant, germinating, and outgrowing bacterial endospores is a valuable prerequisite for gene expression studies, especially in studies on the responses of spores to hostile environmental conditions.

Item URL in elib:https://elib.dlr.de/47246/
Document Type:Article
Title:A Method for Extracting RNA from Dormant and Germinating Bacillus subtilis Strain 168 Endospores
Authors:
AuthorsInstitution or Email of AuthorsAuthors ORCID iD
Moeller, RalfUNSPECIFIEDUNSPECIFIED
Horneck, GerdaUNSPECIFIEDUNSPECIFIED
Rettberg, PetraUNSPECIFIEDUNSPECIFIED
Mollenkopf, H.-J.Microarray Core Facility, MPI-IB–Max Planck Institute for Infection Biology, Berlin, GermanyUNSPECIFIED
Stackebrandt, E.German Collection of Microorganism and Cell Cultures GmbH, Braunschweig, GermanyUNSPECIFIED
Nicholson, W.L.University of Florida, Department of Microbiology and Cell Science, Gainesville, Florida, USA; Kennedy Space Center,UNSPECIFIED
Date:2006
Journal or Publication Title:CURRENT MICROBIOLOGY
Refereed publication:Yes
Open Access:No
Gold Open Access:No
In SCOPUS:No
In ISI Web of Science:Yes
Volume:53
Page Range:pp. 227-231
Publisher:Springer Science+Business Media
Status:Published
Keywords:Bacillus subtilis, RNA,
HGF - Research field:Aeronautics, Space and Transport (old)
HGF - Program:Space (old)
HGF - Program Themes:W FR - Forschung unter Weltraumbedingungen (old)
DLR - Research area:Space
DLR - Program:W FR - Forschung unter Weltraumbedingungen
DLR - Research theme (Project):W - Vorhaben Strahlenbiologie (old)
Location: Köln-Porz
Institutes and Institutions:Institute of Aerospace Medicine > Radiation Biology
Deposited By: Kopp, Kerstin
Deposited On:28 Feb 2007
Last Modified:27 Apr 2009 13:30

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