METHODS AND COMPOSITIONS FOR TARGETED NUCLEIC ACID SEQUENCING
First Claim
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1. A method for preparing a targeted nucleic acid library comprising:
- (a) providing a nucleic acid library comprising a plurality of different nucleic acids and a double-stranded target nucleic acid;
(b) hybridizing a first adapter probe with the target nucleic acid in the presence of a recombinase,wherein the first adapter probe comprises a first portion complementary to a substrate probe and a second portion exogenous to the first portion and complementary to at least a portion of the double-stranded target nucleic acid, andwherein the recombinase promotes hybridizing the second portion with the double-stranded target nucleic acid;
(c) extending the hybridized first adapter probe;
(d) hybridizing the extended adapter probe with the substrate probe, wherein the substrate probe is immobilized on a first substrate; and
(e) extending the hybridized substrate probe to form an anchor probe, thereby obtaining a targeted nucleic acid library.
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Abstract
Embodiments provided herein relate to methods and compositions for obtaining nucleic acid sequence information. Some embodiments provided herein include methods and compositions for preparing nucleic acid libraries. In some embodiments, such nucleic acid libraries are useful for targeted nucleic acid sequencing.
20 Citations
53 Claims
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1. A method for preparing a targeted nucleic acid library comprising:
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(a) providing a nucleic acid library comprising a plurality of different nucleic acids and a double-stranded target nucleic acid; (b) hybridizing a first adapter probe with the target nucleic acid in the presence of a recombinase, wherein the first adapter probe comprises a first portion complementary to a substrate probe and a second portion exogenous to the first portion and complementary to at least a portion of the double-stranded target nucleic acid, and wherein the recombinase promotes hybridizing the second portion with the double-stranded target nucleic acid; (c) extending the hybridized first adapter probe; (d) hybridizing the extended adapter probe with the substrate probe, wherein the substrate probe is immobilized on a first substrate; and (e) extending the hybridized substrate probe to form an anchor probe, thereby obtaining a targeted nucleic acid library. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18)
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19-20. -20. (canceled)
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21. A method of preparing a nucleic acid library comprising:
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(a) providing a plurality of anchor probes; (b) hybridizing a plurality of double-stranded target nucleic acids with the plurality of anchor probes in the presence of a recombinase under conditions wherein the recombinase promotes the hybridizing with the target nucleic acids; (c) hybridizing the plurality of double-stranded target nucleic acids with a plurality of ligation probes in the presence of a recombinase under conditions wherein the recombinase promotes the hybridizing with the target nucleic acids; (d) extending the anchor probes or the ligation probes; and (e) ligating the anchor probes to the ligation probes to form a plurality of different extended ligated probes, thereby preparing the library of nucleic acids.
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22-51. -51. (canceled)
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52. A method of detecting a nucleic acid comprising:
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(a) providing an anchor probe; (b) hybridizing a double-stranded target nucleic acid with the anchor probe in the presence of a recombinase under conditions wherein the recombinase promotes the hybridizing with the target nucleic acid; (c) hybridizing the double-stranded target nucleic acid with a ligation probe in the presence of a recombinase under conditions wherein the recombinase promotes the hybridizing with the target nucleic acid; (d) extending the anchor probe or the ligation probe; (e) ligating the anchor probe to the ligation probe to form an extended ligated probe; and (f) detecting the extended ligated probe, thereby detecting the nucleic acid.
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53-67. -67. (canceled)
Specification