GMNL-464 sequencing by NGS
A total amount of 1 g DNA per sample was used as input material for the DNA sample preparations. Sequencing libraries were generated using NEBNext Ultra DNA Library Prep Kit for Illumina (NEB, USA) following manufacturers recommendations and index codes were added to attribute sequences to each sample. Briefly, the DNA sample was fragmented by sonication to a size of 350 bp, then DNA fragments were end-polished, A-tailed, and ligated with the full-length adaptor for Illumina sequencing with further PCR amplification. At last, PCR products were purified (AMPure XP system) and libraries were analysed for size distribution by Agilent 2100 Bioanalyzer and quantified using real-time PCR. The clustering of the index-coded samples was performed on a cBot Cluster Generation System according to the manufacturers instructions. After cluster generation, the library preparations were sequenced on an Illumina Novaseq 6000 platform and paired-end reads were generated.