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TM3’seq: a tagmentation-mediated 3’ sequencing approach for improving scalability of RNA-seq experiments

Luisa F. Pallares, Serge Picard, Julien F. Ayroles
doi: https://doi.org/10.1101/585810
Luisa F. Pallares
1Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, USA
2Department of Ecology and Evolution, Princeton University, Princeton, USA
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Serge Picard
1Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, USA
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Julien F. Ayroles
1Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, USA
2Department of Ecology and Evolution, Princeton University, Princeton, USA
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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.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted March 22, 2019.
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TM3’seq: a tagmentation-mediated 3’ sequencing approach for improving scalability of RNA-seq experiments
Luisa F. Pallares, Serge Picard, Julien F. Ayroles
bioRxiv 585810; doi: https://doi.org/10.1101/585810
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TM3’seq: a tagmentation-mediated 3’ sequencing approach for improving scalability of RNA-seq experiments
Luisa F. Pallares, Serge Picard, Julien F. Ayroles
bioRxiv 585810; doi: https://doi.org/10.1101/585810

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