Methods and Compositions for Nucleic Acid Purification
AssigneeAffymetrix, Inc.
InventorsGary K. McMaster, Quan N. Nguyen, Franklin R. Witney, Yunqing Ma
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Abstract
Methods of capturing two or more nucleic acids simultaneously from a single sample are provided. Different nucleic acids are captured through cooperative hybridization events on a substrate, or different subsets of particles, or at different selected positions on a spatially addressable solid support. Methods described include enrichment and purification of nucleic acids prior to downstream steps including sequencing of target nucleic acids. Compositions, kits, and systems related to the methods are also described.
Claims20
A method of capturing two or more nucleic acids of interest, the method comprising:
- a) providing a sample comprising or suspected of comprising the nucleic acids of interest;
- b) providing a substrate having associated therewith a plurality of support capture probe;
- c) providing two or more subsets of n target capture probes, wherein n is at least two, wherein each subset of n target capture probes is capable of hybridizing to one of the nucleic acids of interest, and wherein the target capture probes in each subset are capable of hybridizing to one of the support capture probes and thereby associating each subset of n target capture probes with a selected subset of the particles;
- d) contacting the sample, the substrate, and the subsets of n target capture probes;
- and
- e) hybridizing any nucleic acid of interest present in the sample to its corresponding subset of n target capture probes and hybridizing the subset of n target capture probes to its corresponding support capture probe, whereby the hybridizing the nucleic acid of interest to the n target capture probes and the n target capture probes to the corresponding support capture probe captures the nucleic acid on the subset of particles with which the target capture probes are associated,
- wherein the hybridizing the subset of n target capture probes to the corresponding support capture probe is performed at a hybridization temperature which is greater than a melting temperature Tm of a complex between each individual target capture probe and its corresponding support capture probe.
A composition comprising:
- a substrate having associated therewith a plurality of support capture probes;
- and
- two or more subsets of n target capture probes, wherein n is at least two, wherein each subset of n target capture probes is capable of hybridizing to a different nucleic acid of interest, and wherein the target capture probes in each subset are capable of hybridizing to one of the support capture probes and thereby associating each subset of n target capture probes with the substrate,
- wherein when the nucleic acid of interest corresponding to a subset of n target capture probes is present in the composition and is hybridized to the subset of n target capture probes, which target capture probes are hybridized to the corresponding support capture probe, the nucleic acid of interest is hybridized to the subset of n target capture probes at a hybridization temperature which is greater than a melting temperature Tm of a complex between each individual target capture probe and the support capture probe.
Dependencies are inferred from the claim text.