A method for sequencing an unknown flanking sequence at both sides of a known sequence comprises
library construction, flanking sequence capturing and sequencing. The method comprises: employing
a site directed mutagenesis PCR process, designing an appropriate primer on the known sequence (without
mutation sites), taking the constructed
library as the template, screening numerous plasmids in the
library to obtain
a DNA fragment containing the known sequence sites, screening out target sites in the
plasmid library by employing the PCR process, and further obtaining an
insertion sequence 2 Kb-5 Kb and a
plasmid library containing the known sequence, and taking the
plasmid library as the target point to obtain the flanking sequence at the both sides of the known sequence. The provided sequencing method helps to realize
full view acquisition of
genome, helps to solve the technological restriction that researchers cannot describe the
full view of a
gene and consequently cannot precisely research the functions of the
gene when carrying out comprehensive research on the
gene, and is beneficial to improve the efficiency of related researches.