Simplified Multiplexed Genome Sequencing

Based on simplified genome sequencing, the qualified DNA was digested with Mse I restriction endonuclease and a linker with a barcode was added to both sides of the enzyme section. Then, PCR amplification was carried out, and the amplified products were mixed. The required fragments were selected to establish the library. The constructed library was initially quantified using Qubit^®2.0 and diluted to 1 ng·μL⁻¹. The library was examined using an Agilent 2100 Bioanalyzer (Santa Clara, CA, USA). After the length of the inserted fragment was obtained, the effective concentration of the library was accurately quantified using q-PCR (the effective concentration of the library > 2 nmol·L⁻¹) to ensure the quality of the library [12 (link)]. The library was qualified. The pool was mixed according to the effective concentration of different libraries and the amount of data required for the target machine, followed by Illumina Hi-seq PE150 sequencing [13 (link)].