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Quorum Sensing pp 177-192 | Cite as

Global Expression Analysis of Quorum Sensing-Controlled Genes by RNAseq

Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1673)

Abstract

RNA sequencing (RNAseq) enables transcriptional profiling of many organisms. This chapter describes the use of RNAseq in prokaryotes to identify quorum sensing (QS)-controlled transcripts by comparing samples from QS-induced and -uninduced conditions. Briefly, each RNA sample is converted to ds-cDNA in a method that limits amplification of ribosomal RNA species. The ds-cDNA contains adapters that enable sequencing and quantification by next-generation sequencing (NGS).

Key words

RNA sequencing (RNAseq) Next-generation sequencing (NGS) Quorum sensing Not-so-random (NSR) primers Transcriptomics 

Notes

Acknowledgements

A number of people contributed to the methods described in this chapter. Christopher Armour developed the NSR primer approach and methods for library construction [2] and also supported the development of this method for prokaryotic species. Sudha Chugani and Yasuhiro Oda contributed to other aspects of method development, including RNA isolation and library synthesis and analysis. Additionally, this protocol utilizes a number of commercially available kits. Many of the procedures are taken directly from the kit protocols or handbooks.

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Copyright information

© Springer Science+Business Media LLC 2018

Authors and Affiliations

  1. 1.Oregon State UniversityBendUSA

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