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High-throughput RNA isoform sequencing using programmed cDNA concatenation.

Authors :
Al'Khafaji, Aziz M.
Smith, Jonathan T.
Garimella, Kiran V.
Babadi, Mehrtash
Popic, Victoria
Sade-Feldman, Moshe
Gatzen, Michael
Sarkizova, Siranush
Schwartz, Marc A.
Blaum, Emily M.
Day, Allyson
Costello, Maura
Bowers, Tera
Gabriel, Stacey
Banks, Eric
Philippakis, Anthony A.
Boland, Genevieve M.
Blainey, Paul C.
Hacohen, Nir
Source :
Nature Biotechnology; Apr2024, Vol. 42 Issue 4, p582-586, 5p
Publication Year :
2024

Abstract

Full-length RNA-sequencing methods using long-read technologies can capture complete transcript isoforms, but their throughput is limited. We introduce multiplexed arrays isoform sequencing (MAS-ISO-seq), a technique for programmably concatenating complementary DNAs (cDNAs) into molecules optimal for long-read sequencing, increasing the throughput >15-fold to nearly 40 million cDNA reads per run on the Sequel IIe sequencer. When applied to single-cell RNA sequencing of tumor-infiltrating T cells, MAS-ISO-seq demonstrated a 12- to 32-fold increase in the discovery of differentially spliced genes. Programmable concatenation of cDNA molecules increases the throughput of PacBio sequencing about 15-fold. [ABSTRACT FROM AUTHOR]

Details

Language :
English
ISSN :
10870156
Volume :
42
Issue :
4
Database :
Complementary Index
Journal :
Nature Biotechnology
Publication Type :
Academic Journal
Accession number :
176650927
Full Text :
https://doi.org/10.1038/s41587-023-01815-7