Skip Navigation

Publication Detail

Title: TM3'seq: A Tagmentation-Mediated 3' Sequencing Approach for Improving Scalability of RNAseq Experiments.

Authors: Pallares, Luisa F; Picard, Serge; Ayroles, Julien F

Published In G3 (Bethesda), (2020 01 07)

Abstract: RNA-seq has become the standard tool for collecting genome-wide expression data in diverse fields, from quantitative genetics and medical genomics to ecology and developmental biology. However, RNA-seq library preparation is still prohibitive for many laboratories. Recently, the field of single-cell transcriptomics has reduced costs and increased throughput by adopting early barcoding and pooling of individual samples -producing a single final library containing all samples. In contrast, RNA-seq protocols where each sample is processed individually are significantly more expensive and lower throughput than single-cell approaches. Yet, many projects depend on individual library generation to preserve important samples or for follow-up re-sequencing experiments. Improving on currently available RNA-seq methods we have developed TM3'seq, a 3'-enriched library preparation protocol that uses Tn5 transposase and preserves sample identity at each step. TM3'seq is designed for high-throughput processing of individual samples (96 samples in 6h, with only 3h hands-on time) at a fraction of the cost of commercial kits ($1.5 per sample). The protocol was tested in a range of human and Drosophila melanogaster RNA samples, recovering transcriptomes of the same quality and reliability than the commercial NEBNext kit. We expect that the cost- and time-efficient features of TM3'seq make large-scale RNA-seq experiments more permissive for the entire scientific community.

PubMed ID: 31676507 Exiting the NIEHS site

MeSH Terms: 3' Untranslated Regions; Animals; Costs and Cost Analysis; Drosophila melanogaster; Female; Humans; RNA, Messenger/chemistry; RNA, Messenger/genetics; RNA-Seq/economics; RNA-Seq/methods*; RNA-Seq/standards; Reproducibility of Results

Back
to Top