Construction of Metatranscriptomic Libraries for 5' End Sequencing of rRNAs for Microbiome Research

Marja Tiirola; Anita Mäki

doi:10.1007/978-1-0716-1585-0_7

Construction of Metatranscriptomic Libraries for 5' End Sequencing of rRNAs for Microbiome Research

Methods Mol Biol. 2022:2349:137-146. doi: 10.1007/978-1-0716-1585-0_7.

Authors

Marja Tiirola¹, Anita Mäki²

Affiliations

¹ Department of Biological and Environmental Science, University of Jyväskylä, Jyväskylä, Finland. mtiirola@jyu.fi.
² Department of Biological and Environmental Science, University of Jyväskylä, Jyväskylä, Finland.

PMID: 34718994
DOI: 10.1007/978-1-0716-1585-0_7

Abstract

Metatranscriptomic sequencing enables studying community-wide gene expression profiles of microbial samples and getting functional insight on their up- or downregulated pathways. However, shotgun sequencing is not the most efficient way to study expression of ribosomal RNA genes or to compare lot of samples in experimental setups. Here we describe an efficient primer-independent method for processing and barcoding libraries for directional sequencing of the 5' end region of the RNA. When applying size selection of the original RNA, the method forms an optimal solution for the simultaneous analysis of bacterial, archaeal, and eukaryotic rRNA diversity.

Keywords: High-throughput; Metatranscriptomics; Microbiomes; NGS; RNA; SSU rRNA; Sequencing.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Archaea / genetics
Bacteria / genetics
High-Throughput Nucleotide Sequencing
Microbiota* / genetics
RNA, Ribosomal / genetics
RNA, Ribosomal, 16S / genetics

Substances

RNA, Ribosomal
RNA, Ribosomal, 16S