Abstract
Bacterial RNA abundance is monitored to analyze gene expression at the transcriptional level. RNA is typically extracted using organic solvents or commercially available kits. Appropriate techniques for total RNA extraction should be used to harvest sufficient starting material required for further analysis while taking care not to bias sampling of the transcriptome. Here, we describe two methods to obtain high-quality RNA from Yersinia pestis by using TRIzol® Reagent and the PureLink™ RNA Mini Kit. The first method allows us to disrupt and dissolve bacterial cells by using TRIzol® Reagent, followed by phase separation and RNA precipitation. The alternative method using the PureLink™ RNA Mini Kit contains a column purification step. After homogenization, the sample is processed through a membrane to purify RNA.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsSpringer Nature is developing a new tool to find and evaluate Protocols. Learn more
References
Han YP, Xin DS, Song YJ, Ling Z, Bao JY, Tong ZZ, Jin W, Guo ZB, Zhai JH, Du ZM (2004) Microarray analysis of temperature-induced transcriptome of Yersinia pestis. Microbiol Immunol 48(11):791–805
Han Y, Zhou D, Pang X, Zhang L, Song Y, Tong Z, Bao J, Dai E, Wang J, Guo Z (2005) DNA microarray analysis of the heat- and cold-shock stimulons in Yersinia pestis. Microbes Infect 7.0(3):335
Qiu J, Zhou D, Qin L, Han Y, Wang X, Zongmin DU, Song Y, Yang R (2006) Microarray expression profiling of Yersinia pestis in response to chloramphenicol. FEMS Microbiol Lett 263(1):26
Zhou D, Qin L, Han Y, Qiu J, Chen Z, Li B, Song Y, Wang J, Guo Z, Zhai J (2006) Global analysis of iron assimilation and fur regulation in Yersinia pestis. FEMS Microbiol Lett 258(1):9–17
Chomczynski P, Sacchi N (2006) The single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction: twenty-something years on. Nat Protoc 1(2):581–585
Chomczynski P, Sacchi N (1987) Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 162(1):156–159
Wilfinger WW, Mackey K, Chomczynski P (1997) Effect of pH and ionic strength on the spectrophotometric assessment of nucleic acid purity. BioTechniques 22(3):474–476, 478–481
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2018 Springer Nature Singapore Pte Ltd.
About this protocol
Cite this protocol
Wang, H., Han, Y. (2018). Extraction of Total RNA from Yersinia pestis. In: Yang, R. (eds) Yersinia Pestis Protocols. Springer Protocols Handbooks. Springer, Singapore. https://doi.org/10.1007/978-981-10-7947-4_4
Download citation
DOI: https://doi.org/10.1007/978-981-10-7947-4_4
Published:
Publisher Name: Springer, Singapore
Print ISBN: 978-981-10-7946-7
Online ISBN: 978-981-10-7947-4
eBook Packages: Springer Protocols