Massively multiplexed RNA sequencing

US10612088B2 · kind B2 · utility

Assignees

• The Broad Institute, Inc. · US
• Massachusetts Institute of Technology · US

Inventors

• Alexander A. Shishkin · Pasadena, US
• Mitchell Guttman · Cambridge, US
• Christine F. Surka · Pasadena, US
• Eric S. Lander · Cambridge, US

Key dates

• Filing date: Mar 14, 2014
• Grant date: Apr 7, 2020
• Expiry date: Oct 15, 2034

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12Q2563/179
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

A method for parallel sequencing target RNA from samples from multiple sources while maintaining source identification is provided. The method includes providing samples of RNA comprising target RNA from two or more sources; labeling, at the 3′ end, the RNA from the two or more sources with a first nucleic acid adaptor that comprises a nucleic acid sequence that differentiates between the RNA from the two or more sources; reverse transcribing the two or more sources to create a single stranded DNA comprising the nucleic acid sequence that differentiates between the RNA from the two or more sources; amplifying the single stranded DNA to create DNA amplification products that comprise the nucleic acid sequence that differentiates between the RNA from the two or more sources; sequencing the DNA amplification products thereby parallel sequencing target RNA from samples from multiple sources while maintaining source identification.