Abstract
Bacteriophages manipulate bacterial gene expression in order to express their own genes or influence bacterial metabolism. Gene expression can be studied using real-time PCR or microarrays. Either technique requires the prior isolation of high quality RNA uncontaminated by the presence of genomic DNA. We outline the considerations necessary when working with bacteriophage infected bacterial cells. We also give an example of a protocol for extraction and quantification of high quality RNA from infected bacterial cells, using the marine cyanobacterium WH7803 and the phage S-PM2 as a case study. This protocol can be modified to extract RNA from the host/bacteriophage of interest.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Sung, K., et al., A simple and efficient Triton X-100 boiling and chloroform extraction method of RNA isolation from Gram-positive and Gram-negative bacteria. FEMS Microbiology Letterss, 2003. 229(1): 97–101.
Chomczynski, P. and N. Sacchi, Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Analytical Biochemistry, 1987. 162(1): 156–159.
Bustin, S.A., Real-time, fluorescence-based quantitative PCR: a snapshot of current procedures and preferences. Expert review of molecular diagnostics, 2005. 5(4): 493–498.
Casadesus, J., D. Low, and M.A.M.B.R.-S. 2006, Epigenetic gene regulation in the bacterial world. Microbiology and molecular biology reviews, 2006. 70(3): 830–856.
Imbeaud, S., et al., Towards standardization of RNA quality assessment using user-independent classifiers of microcapillary electrophoresis traces. Nucleic Acids Research, 2005. 33: e56 (Online publication).
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2009 Humana Press, a part of Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Shan, J., Clokie, M. (2009). Preparation of RNA from Bacteria Infected with Bacteriophages: A Case Study from the Marine Unicellular Synechococcus sp. WH7803 Infected by Phage S-PM2. In: Clokie, M.R., Kropinski, A.M. (eds) Bacteriophages. Methods in Molecular Biology™, vol 502. Humana Press. https://doi.org/10.1007/978-1-60327-565-1_10
Download citation
DOI: https://doi.org/10.1007/978-1-60327-565-1_10
Publisher Name: Humana Press
Print ISBN: 978-1-60327-564-4
Online ISBN: 978-1-60327-565-1
eBook Packages: Springer Protocols