lcWGS of Sardinops sagax : MTC090722-A1
Genomic DNA was extracted from muscle tissue or fin clips stored in 100% ethanol using Qiagen DNAeasy Blood & Tissue 96 extraction kits (Qiagen, Inc., Valencia, CA). Extracted DNA was run on a 1% agarose gel stained with ethidium bromide to assess DNA quality and was quantified using fluorescence assays on a Perkin-Elmer Victor. 10ng of DNA from each high-quality extraction was plated and allowed to dry at room temperature while covered with a micropore plate cover. DNA was fragmented and tagged (tagmented) with a universal Nextera overhang following the Nextera DNA Library Prep Kit (Illumina, Inc., San Diego, CA) using 1/20th of recommended reagents. Tagmented libraries were amplified with low-cycle PCR and barcoded using Illumina Nextera dual-indices at concentrations of 5uM. Illumina P5 and P7 sequencing primers were added using another round of low-cycle PCR. Tagmented and indexed samples were then normalized (<=25ng) using 96-well SequelPrep Normalization Plates following manufacturer protocol and then pooled for each plate. Pooled libraries were cleaned using AMPure XP beads (Beckman Coulter, Inc., Brea, CA) and eluted in 20ul of TLE buffer. Libraries were sequenced on two lanes of 2 x 150bp paired-end Illumina NovaSeq XB+ at Azenta Genewiz.