Method for detecting guanine-abasic site in DNA
First Claim
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1. A method for detecting the position of a guanine-abasic site generated in a double-stranded DNA, comprising:
- (1) biotinylating an unpaired guanine residue in a double-stranded DNA containing at least one abasic site and the unpaired guanine;
(2) purifying the biotinylated double-stranded DNA on an avidin column;
(3) performing polymerase chain reaction on the purified biotinylated double-stranded DNA, which serves as a template and obtaining an amplification product;
(4) searching for a mutation from guanine to adenine in a base sequence of the amplification product; and
(5) detecting the position of the mutation from guanine to adenine as the position of the guanine-abasic site.
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Abstract
The present invention provides a method for detecting the presence or absence of a guanine-abasic site, the method being a process for detecting guanine opposite at least one abasic sites generated in a double-stranded DNA, comprising:
- (1) step 1 of site-selectively cleaving at least one abasic sites in a double-stranded DNA using an enzyme;
- (2) step 2 of modifying the amino group at position 2 of guanine opposite the abasic sites using a modifier; and
- (3) step 3 of performing polymerase chain reaction on the modified double-stranded DNA obtained by conducting step 1 and step 2, which serves as a template, to search for the presence or absence of an amplification product, the sequence of steps 1 and 2 being not limited to the order presented.
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Citations
7 Claims
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1. A method for detecting the position of a guanine-abasic site generated in a double-stranded DNA, comprising:
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(1) biotinylating an unpaired guanine residue in a double-stranded DNA containing at least one abasic site and the unpaired guanine; (2) purifying the biotinylated double-stranded DNA on an avidin column; (3) performing polymerase chain reaction on the purified biotinylated double-stranded DNA, which serves as a template and obtaining an amplification product; (4) searching for a mutation from guanine to adenine in a base sequence of the amplification product; and (5) detecting the position of the mutation from guanine to adenine as the position of the guanine-abasic site. - View Dependent Claims (2, 3, 4, 5, 6, 7)
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