Enhanced Methods of Ribonucleic Acid Hybridization

摘要

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.

主权项

1. A method of increasing the hybridization efficiency of a probe and a target RNA in a cell lysate, comprising:
heating a cell lysate sample comprising at least one target RNA at a temperature of about 80° C. and 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 cell lysate comprises a cell lysis buffer comprising a chemical denaturant; contacting the cell lysate sample with at least one detectable probe, wherein the at least one probe specifically hybridizes to the target RNA in the cell lysate sample; and detecting hybridization between the probe and the target RNA, wherein the detected hybridization between the probe and the target RNA is increased relative to the hybridization between the probe and the target RNA in the absence of the heating step.