C-PROBE LIBRARIES FOR DNA TARGET ENRICHMENT

摘要

Provided herein is a method for enriching a target nucleic acid molecule. In one embodiment, the method may involve hybridizing a C-probe to a strand of a target nucleic acid to produce a complex, enzymatically removing any 3′ overhanging end from the target nucleic acid of the complex to produce a 3′ hydroxyl group at the 3′ end; extending the 3′ end of the first sequence using the oligonucleotide sequence of the C-probe as a template; enzymatically removing any 5′ overhanging end from the target nucleic acid, either before or after the extending step, to produce an 5′ phosphate group at the end of the second sequence; and ligating the 5′ phosphate group at the end of the second sequence to the 3′ hydroxyl group at the end of the first sequence to produce a circular DNA molecule that contains the target sequence and the complement of the oligonucleotide sequence.

主权项

1. A method of processing a nucleic acid, comprising:
(a) hybridizing a C-probe to a strand of a target nucleic acid to produce a complex, wherein:
(i) said strand comprises a target sequence that is flanked by a first sequence and a second sequence, and(ii) said C-probe comprises a first region that hybridizes to said first sequence, a second region that hybridizes to said second sequence, and an oligonucleotide sequence between said first and second regions, (b) enzymatically removing any 3′ overhang from the target nucleic acid of said complex to produce a 3′ hydroxyl group at the 3′ end of said first sequence; (c) extending the 3′ end of said first sequence using the oligonucleotide sequence of said C-probe as a template, wherein said extending results in a 3′ hydroxyl group that is adjacent to the 5′ end of said second sequence; (d) enzymatically removing any 5′ overhanging from the target nucleic acid, either before or after said extending of step (c), to produce a 5′ phosphate group at the end of said second sequence; and (e) ligating the 5′ phosphate group at the end of the second sequence to the 3′ hydroxyl group that is adjacent to the 5′ end of said second sequence to produce a circular DNA molecule that contains said target sequence and the complement of said oligonucleotide sequence.