Selective Purification of RNA and RNA-Bound Molecular Complexes

摘要

Disclosed are methods for isolating a target nucleic molecule of interest from a sample. The methods include contacting a sample comprising a target nucleic molecule of interest with at least one single stranded nucleic acid targeting probe comprising a nucleic acid sequence between about 30 nucleotide and the length of the target nucleic acid length that hybridizes to the target nucleic molecule of interest under highly stringent hybridization conditions. The probe comprises a capture moiety covalently linked to one or more nucleotide bases in the probe. The targeting probes can be captured with a specific binding agent that specifically binds the capture moiety, thereby isolating the target nucleic molecule of interest. In some embodiments, the sample is further contacted with a crosslinking agent before contacting the sample with a targeting probe. Thus, complexes formed with the target nucleic acid can also be isolated.

主权项

1. A method for isolating a target nucleic acid molecule of interest from a sample, comprising:
contacting a sample comprising a target nucleic acid molecule of interest with at least one single stranded nucleic acid targeting probe that specifically hybridizes to the nucleic acid molecule of interest under highly stringent conditions, wherein the targeting probe comprises a nucleic acid sequence from at least about 30 bases in length to about the length of the target nucleic acid molecule of interest, and wherein the nucleic acid targeting probe further comprises one or more capture moieties covalently linked the targeting probe and optionally a detectable label; contacting the sample with protein-nucleic acid crosslinking agent, a nucleic acid-nucleic acid crosslinking agent, a protein-protein crosslinking agent or any combination thereof before contacting the sample with a targeting probe, thereby isolating any protein and/or nucleic acids crosslinked to the target nucleic molecule of interest; and capturing the at least one targeting probe via the one or more capture moieties, thereby isolating the target nucleic acid molecule of interest specifically bound to the targeting probe.