US 12,071,654 B2
Capture probe-based library normalization
Tricia Zwiefelhofer, San Diego, CA (US); and Jason Nathanson, San Diego, CA (US)
Assigned to SEQUENOM, INC, San Diego, CA (US)
Filed by Sequenom, Inc., San Diego, CA (US)
Filed on Apr. 1, 2020, as Appl. No. 16/837,425.
Claims priority of provisional application 62/827,601, filed on Apr. 1, 2019.
Prior Publication US 2020/0308637 A1, Oct. 1, 2020
Int. Cl. C12Q 1/6837 (2018.01); C12Q 1/6874 (2018.01); C40B 40/06 (2006.01)
CPC C12Q 1/6837 (2013.01) [C12Q 1/6874 (2013.01); C40B 40/06 (2013.01); C12Q 2600/158 (2013.01)] 29 Claims
 
1. A method for normalizing a nucleic acid library comprising:
(a) contacting at least some of single-stranded nucleic acid molecules Obtained from the library with a solution comprising a salt and one or more probes,
wherein each of at least some of the single-stranded nucleic acid molecules comprises an A adaptor sequence and a B adaptor sequence,
wherein the one or more probes each has a sequence that is complementary to the A adaptor sequence or the B adaptor sequence,
wherein the one or more probes hybridize to the single-stranded nucleic acid molecules to form hybridization complexes, and
wherein the one or more probes each is conjugated to a first binding member,
(b) contacting the hybridization complexes with solid supports, each solid support conjugated to a second binding member,
wherein the first binding member binds to the second binding member, thereby causing the solid supports to bind to the hybridization complexes to form solid support-bound hybridization complexes,
(c) collecting the solid support-bound hybridization complexes, and
(d) separating the single-stranded nucleic acid molecules from probes in the hybridization complexes, thereby obtaining a normalized library of single-stranded nucleic acid molecules.