US 11,859,178 B2
Nucleic acid library methods
Caroline Julie Gallant, Stockholm (SE); Marlon Stoeckius, Stockholm (SE); and Katherine Pfeiffer, San Francisco, CA (US)
Assigned to 10x Genomics, Inc., Pleasanton, CA (US)
Filed by 10x Genomics, Inc., Pleasanton, CA (US)
Filed on Feb. 7, 2023, as Appl. No. 18/165,721.
Application 18/165,721 is a continuation of application No. 17/963,749, filed on Oct. 11, 2022, granted, now 11,608,498.
Application 17/963,749 is a continuation of application No. 17/690,628, filed on Mar. 9, 2022, granted, now 11,512,308.
Application 17/690,628 is a continuation of application No. PCT/US2021/035211, filed on Jun. 1, 2021.
Claims priority of provisional application 63/033,577, filed on Jun. 2, 2020.
Prior Publication US 2023/0183684 A1, Jun. 15, 2023
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 15/10 (2006.01); C12Q 1/6837 (2018.01); C12Q 1/6874 (2018.01)
CPC C12N 15/1093 (2013.01) [C12Q 1/6837 (2013.01); C12Q 1/6874 (2013.01)] 30 Claims
 
1. A method for removing all or a portion of a nucleic acid analyte sequence from a double-stranded member of a nucleic acid library, wherein a single-strand of the double-stranded member of the nucleic acid library comprises: a ligation sequence, a barcode, a reverse complement of a first adaptor, a capture domain, a sequence that is complementary to all or a portion of the nucleic acid analyte sequence, and a second adaptor, wherein the method comprises:
(a) ligating ends of the double-stranded member using the ligation sequence to generate a circularized double-stranded nucleic acid;
(b) amplifying the circularized double-stranded nucleic acid using a first primer and a second primer to generate a double-stranded member of the nucleic acid library lacking all or a portion of the nucleic acid analyte sequence, wherein:
the first primer comprises: (i) a sequence substantially complementary to the reverse complement of the first adaptor and (ii) a first functional domain; and
the second primer comprises: (i) a sequence substantially complementary to a sequence from a 5′ region of the nucleic acid analyte sequence, and (ii) a second functional domain.