Efficient and cost-effective bacterial mRNA sequencing from low input samples through ribosomal RNA depletion.

ABSTRACT:

Background

RNA sequencing is a powerful approach to quantify the genome-wide distribution of mRNA molecules in a population to gain deeper understanding of cellular functions and phenotypes. However, unlike eukaryotic cells, mRNA sequencing of bacterial samples is more challenging due to the absence of a poly-A tail that typically enables efficient capture and enrichment of mRNA from the abundant rRNA molecules in a cell. Moreover, bacterial cells frequently contain 100-fold lower quantities of RNA compared to mammalian cells, which further complicates mRNA sequencing from non-cultivable and non-model bacterial species. To overcome these limitations, we report EMBR-seq (Enrichment of mRNA by Blocked rRNA), a method that efficiently depletes 5S, 16S and 23S rRNA using blocking prime

SUBMITTER: Wangsanuwat C 

PROVIDER: S-EPMC7565789 | biostudies-literature | 2020 Oct

REPOSITORIES: biostudies-literature