METHODS AND COMPOSITIONS FOR IMPROVING REMOVAL OF RIBOSOMAL RNA FROM BIOLOGICAL SAMPLES

摘要

The invention generally relates to compositions for maximizing capture of affinity-labeled molecules on solid supports. The disclosed methods and compositions were developed to maximize depletion of ribosomal RNA from total RNA samples, which is useful to improve the quality of RNA preparations used for applications such as massively parallel sequencing. The RNA depletion method is based on using long affinity-labeled RNA molecules that are complementary to all or part of the target ribosomal RNAs, as subtractive hybridization probes.

主权项

1. A method for removing rRNA from an rRNA-containing sample comprising:
obtaining an RNA sample from a biological source, said RNA sample containing rRNA; mixing said RNA sample with one or more RNA subtractive hybridization probe(s) which is/are complementary to one or more rRNA molecules in the RNA sample, wherein the RNA subtractive hybridization probe(s) comprise an affinity label; incubating the RNA sample/RNA subtractive hybridization probe mixture under conditions effective to allow hybridization of the probe(s) with the rRNA in the sample RNA; incubating the hybridized probe rRNA mixture under conditions effective to associate the affinity label in the hybridized probe/rRNA complex with a solid support by linking it to an affinity label recognition molecule attached to the solid support, wherein incubating the hybridized probe rRNA mixture is carried out in the presence of a reaction composition containing components that facilitate association of the hybridized probe/rRNA with the solid support; removing the hybridized probe/sample RNA/solid support complex; and recovering the remaining RNA in the sample.