WGS of Alosa alosa : 10X sequencing
DNA was extracted using a phenol/chloroform protocol and gDNA quality and purity was assessed using spectrophotometry, fluorometry and capillary electrophoresis. 10X Chromium Library was prepared according to 10X Genomics protocols using the Genome Reagent Kits v2. Optimal performance has been characterized on input gDNA with a mean length greater than 50 kb (~156Kb). GEM reactions were performed on 0,625 ng of genomic DNA, and DNA molecules were partitioned and amplified into droplets to introduce a 16-bp partition barcodes. GEM reactions were thermally cycled (30C for 3h and 65C for 10 min; held at 4C) and after amplification the droplets were fractured. P5 and P7 primers, Read 2, and Sample Index were added during library construction. The library was amplified using 10 cycles of PCR and the DNA was subsequently size selected to 450 pb by performing a double purification on AMPure Xp beads. Library quality was assessed using a Fragment Analyzer and quantified by qPCR using the Kapa Library Quantification Kit. Sequencing has been performed on an Illumina HiSeq3000 using a paired-end read length of 2x150 pb with the Illumina HiSeq3000 sequencing kits.
Aalosa_10X