Method for detecting guanine-abasic site in DNA
AssigneeNational Institute of Advanced Industrial Science and Technology
InventorChun Wu
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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.
Claims7
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;
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- (5) detecting the position of the mutation from guanine to adenine as the position of the guanine-abasic site.
Dependencies are inferred from the claim text.