METHOD FOR SYNCHRONOUSLY SEQUENCING SENSE STRAND AND ANTISENSE STRAND OF DNA

摘要

Disclosed is a method for synchronously sequencing a sense strand and an antisense strand of DNA. The method comprises: performing a first round of rolling circle amplification by using a circular DNA molecule to be sequenced as a template to generate a first part of a template strand; hybridizing a blocked or unblocked amplification primer on the first part of the template strand, and then continuing performing a second round of rolling circle amplification by using the circular DNA molecule as the amplification template to generate a second part of the template strand; using either one of the first part of the template strand and the second part of the template strand as a first-strand sequencing template, and performing multiple displacement amplifications by using the other part of the template strand as the amplification template to generate a second-strand sequencing template; and hybridizing a first-strand sequencing primer and a second-strand sequencing primer on the first-strand sequencing template and the second-strand sequencing template respectively, and simultaneously performing first-strand sequencing and second-strand sequencing to obtain the sequences of a sense strand and an antisense strand of DNA. According to the method of the present invention, simultaneous sequencing from both ends of DNA can be realized, thereby greatly saving on sequencing time and sequencing cost, and improving sequencing throughput.