DNA sequencing method and DNA sample preparation method

摘要

A kit and a method of DNA sequencing including digesting a sample DNA with a restriction enzyme to obtain a DNA fragment; introducing an oligonucleotide having a definite base sequence into the DNA fragment at the 3' terminus; and performing a complementary strand extension reaction, using a labeled primer. The complementary strand extension reaction uses as a template, a single strand of the DNA fragment having the oligonucleotide introduced thereinto, to obtain a labeled extended primer having a base sequence complementary to the single strand of the DNA fragment. Next a sequencing reaction with the labeled primer and using the single strand of the DNA fragment having the oligonucleotide introduced thereinto, is performed with the extended labeled primer, using as a template, a part of the single strand of the sample DNA having the base sequence of the single strand of the DNA fragment and a contiguous sequence adjacent thereto, or a single strand of the sample DNA. Finally the products of the sequencing reaction are electrophoresed to determine the base sequence of the DNA fragment and at least a part of the base sequence of the sample DNA adjacent to the base sequence of the DNA fragment.