Methods for depleting RNA from nucleic acid samples

摘要

The invention relates to methods of depleting RNA from a nucleic acid sample. The RNA may be any RNA, including, but not limited to, rRNA, tRNA, and mRNA. The method is useful for depleting RNA from a nucleic acid sample obtained from a fixed paraffin-embedded tissue (FPET) sample. The method may also be used to prepare cDNA, in particular, a cDNA library for further analysis or manipulation.

主权项

1. A method of depleting target RNA molecules in a mixture of nucleic acids comprising target and non-target RNA molecules comprising:
a. contacting a nucleic acid sample with a multiplicity of DNA probes complementary to a target RNA, wherein the multiplicity of DNA probes is capable of specifically hybridizing to the entire full length sequence of the target RNA, includes at least two probes complementary to sequences that overlap one another and forms a DNA-RNA hybrid; and b. contacting the nucleic acid sample with a ribonuclease (RNase) that specifically recognizes the DNA-RNA hybrid; wherein the target RNA in the DNA-RNA hybrid is degraded.