US 12,110,537 B2
Capture reactions
Mark Umbarger, Brookline, MA (US); Gregory Porreca, Cambridge, MA (US); Charles Towne, Shrewsbury, MA (US); and George Church, Brookline, MA (US)
Assigned to Molecular Loop Biosciences, Inc., Woburn, MA (US)
Filed by Molecular Loop Biosciences, Inc., Cambridge, MA (US)
Filed on Jun. 15, 2020, as Appl. No. 16/901,467.
Application 16/901,467 is a continuation of application No. 16/256,574, filed on Jan. 24, 2019, granted, now 10,683,533.
Application 16/256,574 is a continuation of application No. 13/448,961, filed on Apr. 17, 2012, granted, now 10,227,635, issued on Mar. 12, 2019.
Claims priority of provisional application 61/624,778, filed on Apr. 16, 2012.
Prior Publication US 2021/0123094 A1, Apr. 29, 2021
Int. Cl. C12Q 1/6816 (2018.01); C12Q 1/6806 (2018.01); C12Q 1/6813 (2018.01); C12Q 1/6869 (2018.01)
CPC C12Q 1/6816 (2013.01) [C12Q 1/6806 (2013.01); C12Q 1/6813 (2013.01); C12Q 1/6869 (2013.01)] 16 Claims
OG exemplary drawing
 
1. 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.