학술논문
High-throughput RNA isoform sequencing using programmed cDNA concatenation
Document Type
Original Paper
Author
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: The Science and Business of Biotechnology. 42(4):582-586
Subject
Language
English
ISSN
1087-0156
1546-1696
1546-1696
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.
Programmable concatenation of cDNA molecules increases the throughput of PacBio sequencing about 15-fold.