1. High-throughput RNA isoform sequencing using programmed cDNA concatenation.
- 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., and Hacohen, Nir
- 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]
- Published
- 2024
- Full Text
- View/download PDF