AB1_R1041_NGS
Library preparation and sequencing were performed by the Falcon automation system to minimize human error and significantly improve data reliability and consistency. The genomic DNA was fragmented by Frag enzyme to a size of 350bp, then DNA fragments will be end-polished, A-tailed, and ligated with the adaptor for DNB sequencing with further PCR amplification. The PCR product was heat-denatured together with a special molecule that was reverse-complemented to 1 special strand of the PCR product, and the single-strand molecule will be ligated using DNA ligase. The remaining linear molecule was digested with the exonuclease, finally obtaining a single-strand circular DNA library. The library was checked with Qubit and real-time PCR for quantification and a bioanalyzer for size distribution detection. Quantified libraries will be pooled in equal amounts to make DNA Nanoballs (DNB) and sequenced on a DNBseq-T7 platform using PE150 read length. 1 G of raw data per sample was generated.