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.
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