Method for detecting guanine-abasic site in dna
AssigneeNational Institute of Advanced Industrial Science and Technology
InventorChun WU
Life of this patent
Estimating the term…
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.
Claims8
A method for detecting the presence or absence of a guanine-abasic site, the method being a process for detecting guanines 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.
A method for quantifying 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 real-time PCR on the modified double-stranded DNA obtained by conducting step 1 and step 2, which serves as a template, to quantify an amplification product, the sequence of steps 1 and 2 being not limited to the order presented.
A kit for detecting the presence or absence of a guanine-abasic site by performing polymerase chain reaction on a double-stranded DNA, which serves as a template, to search for the presence or absence of an amplification product, the kit comprising an endonuclease for site-selectively cleaving at least one abasic sites in a double-stranded DNA and glyoxal or a derivative of glyoxal for modifying the amino group at position 2 of guanine.
A method for detecting the position 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) a step of biotinylating an unpaired guanine residue in a double-stranded DNA containing at least one abasic sites and the unpaired guanine;
- (2) a step of purifying the biotinylated double-stranded DNA on an avidin column;
- and
- (3) a step of performing polymerase chain reaction on the purified biotinylated double-stranded DNA, which serves as a template, to search for the presence or absence of a mutation in a base sequence of an amplification product.
A kit for detecting the position of a guanine-abasic site, comprising
- a reagent for biotinylating at least one unpaired guanine residues in a double-stranded DNA containing at least one abasic sites and the unpaired guanine
- an avidin column, and
- a reagent for performing polymerase chain reaction.
Dependencies are inferred from the claim text.