A BAC end sequencing method

A BAC end sequencing method is disclosed. The method includes following steps of 1) connecting a target segment to a cloning vector to obtain a recombinant vector, 2) subjecting the recombinant vector to digestion with a restriction enzyme, recovering linear DNA segments longer than the cloning vector to obtain a DNA segment containing two ends of the target segment, wherein the target segment at least comprises two digestion recognition sites of the restriction enzyme and the cloning vector does not have digestion recognition sites of the restriction enzyme, and 3) connecting the DNA segment containing two ends of the target segment to achieve cyclization so as to obtain a plasmid containing the ends of the target segment, and sequencing the plasmid, thus achieving end sequencing of the target segment. Experiments show that BAC end sequencing by the method is 100% successful, and therefore the problem that the success rate of BAC end sequencing is low is overcome, BAC end sequencing is made simple and feasible and is low in cost, and the method is particularly suitable for large-scale BAC end sequencing.