METHOD FOR MEASURING TARGET DNA
The present invention provides a method for adequately analyzing a target DNA after genomic DNA cleavage reaction with a restriction enzyme. More specifically, the present invention provides a method for measuring a target DNA, including (1) cleaving a genomic DNA with a restriction enzyme in a solution to obtain a mixed solution containing (a) completely cleaved DNA fragments consisting of the target DNA, (b) incompletely cleaved DNA fragments comprising the target DNA, and (c) DNA fragments not comprising the target DNA; (2) removing the DNA fragments of (b) from said mixed solution to obtain a solution containing the DNA fragment of (a) and the DNA fragments of (c); and (3) analyzing the target DNA in the solution obtained in (2).
1 : A method for measuring a target DNA, comprising:
(1) cleaving a genomic DNA with a restriction enzyme in a solution to obtain a mixed solution comprising (a) a completely cleaved DNA fragment consisting of the target DNA, (b) incompletely cleaved DNA fragments comprising the target DNA, and (c) DNA fragments not comprising the target DNA;
(2) removing (b) the incompletely cleaved DNA fragments comprising the target DNA from said mixed solution to obtain a solution comprising (a) the completely cleaved DNA fragment consisting of the target DNA and (c) DNA fragments not comprising the target DNA; and
(3) analyzing the target DNA in said solution obtained in (2).
2 : The method according to claim 1 , wherein said removal is performed using a nucleic acid probe specific for non-target DNA.
3 : The method according to claim 2 , wherein the nucleic acid probe specific for the non-target DNA is a nucleic acid probe (1), (2) or (3):
(1) a nucleic acid probe specific for 5′-flanking DNA;
(2) a nucleic acid probe specific for 3′-flanking DNA; or
(3) a combination of the nucleic acid probe specific for the 5′-flanking DNA and the nucleic acid probe specific for the 3′-flanking DNA.
4 : The method according to claim 1 , wherein said analysis is performed using a nucleic acid probe specific for the target DNA.
5 : The method according to claim 1 , wherein a modified nucleobase in the target DNA is analyzed.
6 : The method according to claim 5 , wherein the modified nucleobase is methylcytosine.
7 : The method according to claim 1 , wherein said analysis is performed by immunoassay, mass spectrometry, electrochemical analysis, high performance liquid chromatography analysis, or nanopore analysis.