NGS-DNA-Seq of Phthonandria atrilineata: adult mussle
Genomic DNA was extracted from the muscle tissue of adult Phthonandria atrilineata specimens using a standard phenol-chloroform extraction method. The quality and quantity of the extracted DNA were assessed using Nanodrop spectrophotometry and Qubit fluorometry. For whole genome sequencing (WGS), a genomic DNA library was prepared using a PCR-based method. The DNA was first fragmented to an average size of 350 bp using a Covaris S220 ultrasonicator. The fragmented DNA was then end-repaired, A-tailed, and ligated with Illumina-compatible adapters. The adapter-ligated DNA fragments underwent size selection to obtain the desired insert size range (300-400 bp) using AMPure XP beads. The size-selected library was then amplified using random PCR with Illumina-specific primers to enrich for properly ligated fragments and to add index sequences for multiplexing. The final library quality was assessed using an Agilent 2100 Bioanalyzer, and the quantity was determined by qPCR. The prepared library was then sequenced on an Illumina NovaSeq 6000 platform using paired-end 150 bp sequencing chemistry, following the manufacturer's protocols.
SUB14765118
PHAT_NGS_2024