Methods and compositions for depleting abundant rna transcripts
AssigneeApplera Corporation
InventorsLeopoldo Mendoza, Sharmili Moturi, JR., Robert Setterquist, John Penn Whitley
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Abstract
The present invention concerns a system for isolating, depleting, and/or preventing the amplification of a targeted nucleic acid, such as mRNA or rRNA, from a sample comprising targeted and nontargeted nucleic acids.
Claims31
A method of depleting hemoglobin mRNA in a hemoglobin mRNA-containing sample comprising:
- binding a mixture of capture nucleic acids of SEQ ID NO:19-SEQ ID NO:28 to the sample in a reaction mixture;
- and
- removing hemoglobin mRNA bound to the mixture of capture nucleic acids from the reaction mixture.
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A kit, in a suitable container, comprising a mixture of capture nucleic acids of SEQ ID NO:19-SEQ ID NO:28 and a super-paramagnetic bead.
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A method of preventing poly(dT) primed reverse transcription of a target hemoglobin mRNA in a hemoglobin mRNA-containing sample comprising:
- binding to the sample a first primer mix comprising SEQ ID NO:63-SEQ ID NO:73 that is specific to the target hemoglobin mRNA;
- binding to the sample a second primer comprising a poly(dT) sequence;
- and
- reverse transcribing the RNA in the hemoglobin mRNA-containing sample to form cDNA;
- wherein the first primer mix prevents the reverse transcription of the target hemoglobin mRNA by the second primer.
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Dependencies are inferred from the claim text.