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US 12,110,537 B2Granted
Capture reactions
AssigneeMolecular Loop Biosciences, Inc.
InventorsMark Umbarger, Gregory Porreca, Charles Towne, George Church
Life of this patent
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Abstract
The invention generally relates to methods of performing a capture reaction. In certain embodiments, the method involves obtaining a nucleic acid, fragmenting the nucleic acid, and capturing a target sequence on the nucleic acid fragment using a capture moiety, such as a molecular inversion probe.
Claims16
A method of improving performance of molecular inversion probe capture reactions, the method comprising:
- fragmenting nucleic acid into a plurality of fragments by heating a buffer comprising the nucleic acid to about 65 degrees C. for at least about 16 hours to shift pH of the buffer to between about 6 and about 6.5 to hereby initiate acid hydrolysis;
- introducing a plurality of molecular inversion probes (MIPs) to the plurality of fragments, wherein each of the plurality of MIPs hybridize to the fragments and captures a different one of a plurality of overlapping subregions of the same strand of a fragment, wherein the fragmenting step promotes hybridization of the MIPs to the fragments;
- circularizing the hybridized MIPs;
- amplifying the circularized MIPs to generate amplicons;
- and
- sequencing the amplicons.
3 more elements
A method of improving performance of molecular inversion probe capture reactions, the method comprising:
- fragmenting genomic nucleic acid suspended in a buffer into a plurality of fragments wherein the fragmenting is performed by heating the buffer to about 65 degrees C. for at least about 16 hours to thereby lower pH and initiate acid hydrolysis;
- introducing a plurality of molecular inversion probes (MIPs) to the plurality of fragments, wherein each of the plurality of MIPs hybridize to the fragments and captures a different one of a plurality of overlapping subregions of the same strand of a fragment, wherein the fragmenting step promotes hybridization of the MIPs to the plurality of fragments;
- circularizing the hybridized MIPs;
- amplifying the circularized MIPs to generate amplicons;
- and
- sequencing the amplicons.
3 more elements
Dependencies are inferred from the claim text.