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Preparation of Vibrio cholerae Samples for RNA-seq Analysis
1Department of Medical Microbiology and Immunology, University of Toledo College of Medicine and Life Sciences, Toledo, OH, USA. Jyl.Matson@utoledo.edu.
Methods in Molecular Biology (Clifton, N.J.)
|July 27, 2018
Summary
Massively parallel cDNA sequencing, or RNA-seq, offers a sensitive, unbiased method for analyzing gene expression. This study details the RNA isolation and cDNA library preparation for V. cholerae transcriptomic analysis using Illumina technology.
Area of Science:
- Molecular Biology
- Genomics
- Bioinformatics
Background:
- RNA sequencing (RNA-seq) provides an unbiased assessment of transcript abundance.
- RNA-seq offers higher resolution and sensitivity than microarrays.
- This technique can identify rare transcripts and small RNAs (sRNAs).
Purpose of the Study:
- To describe sample preparation for transcriptomic analysis of V. cholerae.
- To detail the process of generating barcoded cDNA libraries for RNA-seq.
Main Methods:
- RNA isolation from V. cholerae.
- cDNA synthesis and library preparation.
- Illumina sequencing technology-based library generation.
Main Results:
- A comprehensive protocol for V. cholerae RNA-seq library preparation was established.
- Barcoded cDNA libraries were generated, ready for high-throughput sequencing.
Conclusions:
- The described method facilitates robust transcriptomic analysis of V. cholerae.
- This protocol enables sensitive detection and quantification of transcripts, including sRNAs.
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