Use of Single-Stranded Nucleic Acid Binding Proteins In Sequencing
First Claim
Patent Images
1. A method for stabilizing a nucleic acid sequencing reaction, the method comprising the steps of:
- exposing a mixture comprising a template, a polymerase, a primer, and at least one nucleotide to a single-stranded nucleic acid binding protein;
wherein said single-stranded nucleic acid binding protein binds to said template.
8 Assignments
0 Petitions
Accused Products
Abstract
The invention provides methods for stabilizing a nucleic acid sequencing reaction. Generally, methods of the invention include exposing a target nucleic acid to a single-stranded nucleic acid binding protein and performing a sequencing reaction.
-
Citations
45 Claims
-
1. A method for stabilizing a nucleic acid sequencing reaction, the method comprising the steps of:
-
exposing a mixture comprising a template, a polymerase, a primer, and at least one nucleotide to a single-stranded nucleic acid binding protein;
wherein said single-stranded nucleic acid binding protein binds to said template. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11)
-
-
12. A method for sequencing a polynucleotide, the method comprising the steps of:
-
(a) stabilizing a nucleic acid template/primer complex with a single-stranded nucleic acid binding protein;
(b) exposing said complex to a polymerase and at least one nucleotide capable of extending said primer;
(c) determining whether said nucleotide extends said primer;
(d) repeating said exposing and determining steps; and
(e) compiling a sequence of said polynucleotide based upon an order of nucleotides added to said primer. - View Dependent Claims (13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24)
-
-
25. A method for sequencing a nucleic acid template, the method comprising the steps of:
-
(a) exposing a nucleic acid template to a labeled nucleotide, a polymerase and a single-stranded nucleic acid binding protein under conditions that allow incorporation of said nucleotide into a primer attached to said template, wherein said single-stranded nucleic acid binding protein increases fidelity of said polymerase upon binding of said protein to said template;
(b) detecting incorporation of said nucleotide into said primer;
(c) repeating steps (a) and (b) at least once; and
(d) compiling a sequence of said template based upon an order of incorporated nucleotides. - View Dependent Claims (26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 39)
-
-
37. A method for sequencing a nucleic acid template, the method comprising the steps of:
-
(a) exposing a nucleic acid template to a nucleotide, a polymerase and a single-stranded nucleic acid binding protein under conditions that allow incorporation of said nucleotide into a primer attached to said template, said nucleotide comprising a first label, and said single-stranded nucleic acid binding protein comprising a second label, wherein one of said first and second labels comprises a donor fluorophore and the other of said labels comprises an acceptor fluorophore; and
wherein, upon binding of said single-stranded nucleic acid binding protein to said template and incorporation of said nucleotide analogue into said primer, said acceptor fluorophore is optically detectable;
(b) detecting said acceptor fluorophore, thereby to detect incorporation of said nucleotide into said primer;
(c) repeating steps (a) and (b) at least once; and
(d) compiling a sequence of said template based upon an order of incorporated nucleotides. - View Dependent Claims (38, 40, 41)
-
-
42. A method for sequencing a nucleic acid template, the method comprising the steps of:
-
(a) exposing a nucleic acid template to a labeled nucleotide and a polymerase under conditions that allow incorporation of said nucleotide into a primer attached to said template, said template being attached to a substrate-bound single-stranded nucleic acid binding protein such that said template is individually optically resolvable;
(b) detecting incorporation of said nucleotide into said primer;
(c) repeating steps (a) and (b) at least once; and
(d) compiling a sequence of said template based upon an order of incorporated nucleotides. - View Dependent Claims (43, 44, 45)
-
Specification