METHOD FOR AMPLIFICATION OF DNA FRAGMENT

摘要

To measure the methylation degree of a genomic DNA simply and correctly as well as exhaustively, there is provided a method for amplifying a DNA fragment, characterized by comprising the following steps (1) to (5):
(1) digesting a sample double-stranded DNA with a first restriction enzyme;
(2) adding a double-stranded DNA fragment consisting of a first oligonucleotide and a second oligonucleotide to a restriction enzyme cleavage end of the double-stranded DNA fragment produced in step (1) to obtain a DNA fragment;
(3) digesting the DNA fragment obtained in step (2) with a second restriction enzyme;
(4) allowing the double-stranded DNA fragment produced in step (3) to coexist with a third oligonucleotide and a fourth oligonucleotide in the presence of a ligase; and
(5) heating the double-stranded DNA fragment obtained in step (4), thereby dissociating the double-stranded DNA fragment and then performing a polymerase chain reaction to selectively amplify a DNA fragment having the cleavage end of the first restriction enzyme and the cleavage end of the second restriction enzyme.