Method, primer and kit for determining base type
First Claim
1. A base type determination method for determining a base type of a monobasic substituted region of a target nucleic acid, the method comprising the steps of:
- (a) preparing a solution containing a target double-stranded nucleic acid having the monobasic substituted region, a base type determination primer, a DNA polymerase, and dNTPs;
(b) causing the base type determination primer to hybridize to the target double-stranded nucleic acid in the solution, and causing a primer extension reaction to start progressing from the base type determination primer; and
(c) analyzing the degree of progress of the primer extension reaction to determine the base type of the substituted region, wherein the base type determination primer consists of a first single-stranded nucleic acid which is capable of, when hybridizing to the target double-stranded nucleic acid, hybridizing to one of two strands of the target double-stranded nucleic acid such that a 3′
terminal of the primer corresponds to the substituted region of said one strand of the target double-stranded nucleic acid, and wherein the first single-stranded nucleic acid consists of;
a substitution corresponding region which is located at the 3′
terminal and consists of one base complementary to any one of predictable types of bases in the substituted region of said one strand of the target double-stranded nucleic acid;
an uncomplementary region which is located adjacent to the substitution corresponding region on a 5′
terminal side thereof and consists of two bases uncomplementary to said one strand of the target double-stranded nucleic acid; and
a complementary region which is located adjacent to the uncomplementary region on the 5′
terminal side thereof and consists of five or more bases complementary to said one strand of the target double-stranded nucleic acid.
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Accused Products
Abstract
A base type determination primer for determining a base type of a monobasic substituted region of a target nucleic acid, wherein the primer consists of a single-stranded nucleic acid which is capable of hybridizing to the target nucleic acid such that a 3′ terminal thereof corresponds to the substituted region of the target nucleic acid, and wherein the single-stranded nucleic acid includes: a substitution corresponding region located at the 3′ terminal and consisting of a base complementary to any one of predictable types of bases in the substituted region of the target nucleic acid; an uncomplementary region adjacent to the substitution corresponding region on the 5′ terminal side thereof and consisting of two bases uncomplementary to the target nucleic acid; and a complementary region adjacent to the uncomplementary region on the 5′ terminal side thereof and consisting of five or more bases complementary to the target nucleic acid.
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Citations
36 Claims
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1. A base type determination method for determining a base type of a monobasic substituted region of a target nucleic acid, the method comprising the steps of:
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(a) preparing a solution containing a target double-stranded nucleic acid having the monobasic substituted region, a base type determination primer, a DNA polymerase, and dNTPs;
(b) causing the base type determination primer to hybridize to the target double-stranded nucleic acid in the solution, and causing a primer extension reaction to start progressing from the base type determination primer; and
(c) analyzing the degree of progress of the primer extension reaction to determine the base type of the substituted region, wherein the base type determination primer consists of a first single-stranded nucleic acid which is capable of, when hybridizing to the target double-stranded nucleic acid, hybridizing to one of two strands of the target double-stranded nucleic acid such that a 3′
terminal of the primer corresponds to the substituted region of said one strand of the target double-stranded nucleic acid, andwherein the first single-stranded nucleic acid consists of;
a substitution corresponding region which is located at the 3′
terminal and consists of one base complementary to any one of predictable types of bases in the substituted region of said one strand of the target double-stranded nucleic acid;
an uncomplementary region which is located adjacent to the substitution corresponding region on a 5′
terminal side thereof and consists of two bases uncomplementary to said one strand of the target double-stranded nucleic acid; and
a complementary region which is located adjacent to the uncomplementary region on the 5′
terminal side thereof and consists of five or more bases complementary to said one strand of the target double-stranded nucleic acid. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22)
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23. A base type determination primer for determining a base type of a monobasic substituted region of a target nucleic acid,
wherein the primer consists of a single-stranded nucleic acid which is capable of hybridizing to the target nucleic acid such that a 3′ - terminal of the primer corresponds to the substituted region of the target nucleic acid, and
wherein the single-stranded nucleic acid includes;
a substitution corresponding region which is located at the 3′
terminal and consists of one base complementary to any one of predictable types of bases in the substituted region of the target nucleic acid;
an uncomplementary region which is located adjacent to the substitution corresponding region on a 5′
terminal side thereof and consists of two bases uncomplementary to the target nucleic acid; and
a complementary region which is located adjacent to the uncomplementary region on the 5′
terminal side thereof and consists of five or more bases complementary to the target nucleic acid.
- terminal of the primer corresponds to the substituted region of the target nucleic acid, and
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24. A base type determination reagent kit for determining a base type of a monobasic substituted region of a target nucleic acid, the kit comprising a base type determination primer, a DNA polymerase, and dNTPs,
wherein the primer consists of a first single-stranded nucleic acid which is capable of hybridizing to the target nucleic acid such that a 3′ - terminal of the primer corresponds to the substituted region of the target nucleic acid, and
wherein the first single-stranded nucleic acid includes;
a substitution corresponding region which is located at the 3′
terminal and consists of one base complementary to any one of predictable types of bases in the substituted region of the target nucleic acid;
an uncomplementary region which is located adjacent to the substitution corresponding region on a 5′
terminal side thereof and consists of two bases uncomplementary to the target nucleic acid; and
a complementary region which is located adjacent to the uncomplementary region on the 5′
terminal side thereof and consists of five or more bases complementary to the target nucleic acid. - View Dependent Claims (25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36)
- terminal of the primer corresponds to the substituted region of the target nucleic acid, and
Specification