Pool-Sequencing library of Acipenser baerii phenotypic males
Pool-sequencing libraries were prepared using the Illumina TruSeq Nano DNA HT Library Prep Kit (Illumina, San Diego, CA) according to the manufacturer's protocol. After the fragmentation of each gDNA pool (200 ng/pool) by sonication using an M220 Focused-ultrasonicator (COVARIS), the size selection was performed using SPB beads retaining fragments of 550 bp. Following the 3' ends of blunt fragments mono-adenylation and the ligation to specific paired-end adaptors, the amplification of the construction was performed using Illumina-specific primers. Library quality was verified with a Fragment Analyzer (Advanced Analytical Technologies) and then quantified by qPCR using the Kapa Library Quantification Kit (Roche Diagnostics Corp, Indianapolis, IN). The enriched male and female pool libraries were then sequenced using a paired-end multiplexed sequencing mode on a NovaSeq S4 lane (Illumina, San Diego, CA), combining the two pools on the same lane and producing 2 x 150 nt with Illumina NovaSeq Reagent Kits according to the manufacturer's instructions.