Methods for making libraries for nucleic acid sequencing
AssigneeBecton, Dickinson and Company
InventorAlvaro GODINEZ
Life of this patent
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Abstract
Disclosed herein include methods, compositions, and kits suitable for use in generating libraries for nucleic acid sequencing. There are provided, in some embodiments, a plurality of protein complexes. Each protein complex can comprise a transposome and a programmable DNA binding unit capable of specifically binding to a user-selected binding site on a target double-stranded DNA (dsDNA). The binding site for each of the plurality of protein complexes can be different from each other. The transposome can comprise a transposase, a first adaptor, and a second adaptor. The first adaptor, the second adaptor, or both, can be a sequencing adaptor.
Claims44
A composition, comprising a plurality of protein complexes, wherein each of the plurality of protein complexes comprises a transposome and a programmable DNA binding unit capable of specifically binding to a binding site on a target double-stranded DNA (dsDNA), wherein the transposome comprises a transposase, a first adaptor and a second adaptor, and wherein the binding site for each of the plurality of protein complexes is different from each other.
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A reaction mixture, comprising
- a composition comprising a plurality of protein complexes, wherein each of the plurality of protein complexes comprises a transposome and a programmable DNA binding unit capable of specifically binding to a binding site on a target double-stranded DNA (dsDNA), wherein the transposome comprises a transposase, a first adaptor and a second adaptor, and wherein the binding site for each of the plurality of protein complexes is different from each other;
- and
- sample nucleic acids suspected of comprising one or more target dsDNA.
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A method for generating a sequencing library, comprising:
- contacting a composition with a sample suspected of comprising a plurality of target double-stranded DNA (dsDNA) to form a reaction mixture, wherein the composition comprises a plurality of protein complexes, wherein each of the plurality of protein complexes comprises a transposome and a programmable DNA binding unit capable of specifically binding to a binding site on a target double-stranded DNA (dsDNA), wherein the transposome comprises a transposase, a first adaptor and a second adaptor, and wherein the binding site for each of the plurality of protein complexes is different from each other;
- incubating the reaction mixture to generate a plurality of dsDNA fragments each comprising the first adaptor and the second adaptor of one of the plurality of protein complexes at each terminus respectively;
- and
- amplifying the plurality of dsDNA fragments with primers capable of binding to the adaptors at the termini of the dsDNA fragments to generate a sequencing library.
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Dependencies are inferred from the claim text.