Enhanced methods of ribonucleic acid hybridization

US10253353B2 · kind B2 · utility

Assignees

• The Broad Institute, Inc. · US
• The General Hospital Corporation · US

Inventors

• Roby Bhattacharyya · Cambridge, US
• Deborah Hung · Cambridge, US
• Milesh Patel · Jersey City, US

Key dates

• Filing date: Dec 5, 2014
• Grant date: Apr 9, 2019
• Expiry date: Dec 5, 2034

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12Q1/6832
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

This disclosure relates to a method for increasing the hybridization efficiency of a probe and a target RNA in a sample, for example to identify a particular RNA present in the sample. The method includes heating a lysate sample comprising at least one target RNA, such as a tRNA, mRNA or rRNA, at a temperature of about 95° C. for a time sufficient to interfere with secondary structure of the RNA, wherein the time is short enough, such that the RNA in the cell lysate sample are not significantly degraded, and wherein the lysate comprises a cell lysis buffer comprising a chemical denaturant. To detect a target RNA in the lysate, the lysate is contacted with at least one detectable probe, such as a labeled probe, designed to specifically hybridize to the target RNA in the lysate.