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US 11,795,492 B2Granted
Methods of nucleic acid sample preparation
AssigneeArcherDX, Inc.
InventorsJoshua Stahl, Jason Myers, Brady Culver, Brian Kudlow
Life of this patent
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Abstract
Aspects of the technology disclosed herein relate to methods of preparing and analyzing nucleic acids. In some embodiments, methods for preparing nucleic acids for sequence analysis (e.g., using next-generation sequencing) are provided herein.
Claims22
A method of preparing nucleic acids for analysis, the method comprising:
- (a) adding one or more nucleotides to a 3′ end of a double-stranded nucleic acid comprising a target nucleotide sequence, wherein at least one of the one or more nucleotides is a capture moiety modified nucleotide;
- (b) ligating an adapter nucleic acid to at least one strand of the double-stranded nucleic acid to produce a ligation product comprising the one or more capture moiety modified nucleotides;
- and
- (c) capturing the ligation product by contacting the ligation product with a binding partner of the capture moiety modified nucleotide.
1 more element
A method of preparing nucleic acids for analysis, the method comprising:
- (a) adding one or more nucleotides to a 3′ end of each of a plurality of double-stranded nucleic acids comprising a plurality of different target nucleotide sequences, wherein at least one of the one or more nucleotides is a capture moiety modified nucleotide;
- (b) ligating an adapter nucleic acid to at least one strand of each of the plurality of double-stranded nucleic acids to produce a plurality of ligation products comprising the one or more capture moiety modified nucleotides;
- (c) capturing the plurality of ligation products by contacting the plurality of ligation products with a binding partner of the capture moiety modified nucleotide;
- and
- (d) amplifying the plurality of ligation products, thereby providing a plurality of amplicons.
2 more elements
A method of preparing nucleic acids for analysis, the method comprising:
- (a) adding one or more nucleotides to a 3′ end of a double-stranded nucleic acid comprising a target nucleotide sequence, wherein at least one of the one or more nucleotides is a capture moiety modified nucleotide;
- (b) ligating an adapter nucleic acid to at least one strand of the double-stranded nucleic acid to produce a ligation product comprising the one or more capture moiety modified nucleotides;
- (c) capturing the ligation product by contacting the ligation product with a binding partner of the capture moiety modified nucleotide, thereby providing an immobilized ligation product;
- (d) optionally washing the immobilized ligation product;
- (e) separating the ligation product from the binding partner;
- and
- (f) amplifying the separated ligation product of (e) by a polymerase chain reaction using a first target-specific primer that specifically anneals to the target nucleotide sequence and a first adapter primer that specifically anneals to a complementary sequence of the adapter nucleic acid.
4 more elements
A method of preparing nucleic acids for analysis, the method comprising:
- (a) combining a double-stranded nucleic acid, a modified nucleotide comprising a capture moiety, and an enzyme under conditions in which the enzyme adds the modified nucleotide to a 3′ end of the double-stranded nucleic acid;
- (b) combining the double-stranded nucleic acid, an adapter nucleic acid, and a ligase under conditions in which the ligase ligates the adapter nucleic acid to the double-stranded nucleic acid, thereby producing a ligation product comprising the capture moiety;
- and
- (c) combining the ligation product and a binding partner of the capture moiety under conditions in which the binding partner forms a complex with the ligation product.
1 more element
Dependencies are inferred from the claim text.