Infinite f nano plate reader

Before library preparation, the DNA was quantified by Tecan Infinite F Nano + Plate Reader (Tecan, Maennedorf, Switzerland) using Quant-iT dsDNA BR Assay Kit (Thermo Fisher Scientific, Waltham, MA, USA). The enzymatic fragmentation of DNA and library construction was conducted by Tecan DreamPrep NGS using Celero EZ DNA -seq Core Module Kit (Tecan). The fragmented DNA was amplified using PCR. Short and large DNA fragments were removed using double-sided magnetic bead size selection (AMPure XP, Beckman Coulter, Indianapolis, Indiana, USA). Adapter sequences from Celero 96-Plex Adaptor Plate (Tecan) were added to each sample during library construction. The final concentration for each library was quantified by Tecan Infinite F Nano + Plate Reader using NuQuant NGS Library Quantification Module and Qubit (Tecan). The final fragment distribution was evaluated using a Fragment Analyzer 5200 (Agilent, Santa Clara, CA, USA). Qubit and TapeStation were used to determine the concentration of 10% of the final library before sequencing. The library was sequenced using 2 × 150 bp paired-end sequencing on an Illumina platform (Rome, Italy).