PT  - JOURNAL ARTICLE
AU  - Luisa F. Pallares
AU  - Serge Picard
AU  - Julien F. Ayroles
TI  - TM3’seq: a tagmentation-mediated 3’ sequencing approach for improving scalability of RNA-seq experiments
AID  - 10.1101/585810
DP  - 2019 Jan 01
TA  - bioRxiv
PG  - 585810
4099  - http://biorxiv.org/content/early/2019/03/22/585810.short
4100  - http://biorxiv.org/content/early/2019/03/22/585810.full
AB  - 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.