WGS of Alosa alosa : ONT sequencing
DNA was extracted using a phenol/chloroform protocol and gDNA quality and purity was assessed using spectrophotometry, fluorometry and capillary electrophoresis. Additional purification steps were performed using AMPure XP beads (Beckman Coulter). All libraries preparations and sequencing were performed using Oxford Nanopore Ligation Sequencing Kits SQK-LSK108 or SQK-LSK109 according to the manufacturers instructions. For the SQK-LSK108 sequencing Kit, 50g of DNA was purified then sheared to 20kb using the megaruptor1 system (Diagenode). For each library, a DNA-damage repair step was performed on 5g of DNA. Then an END-repair-dA-tail step was performed for adapters ligation. Libraries were loaded onto 8 R9.4 flowcells and sequenced on a GridION instrument at a concentration of 0.1pM for 48 h. For the SQK-LSK109 sequencing Kit, 30g of DNA was purified then sheared to 10kb using the megaruptor1 system (Diagenode). For each library, a one step DNA-damage repair + END-repair-dA-tail procedure was performed on 2g of DNA. Adapters were then ligated to DNAs in the library. Libraries were loaded onto 2 R9.4.1 flowcells and sequenced on a GridION instrument at a concentration of 0.06pM for 48h and onto 1 R9.4.1 flowcell and sequenced on a PromethION instrument at a concentration of 0,04pM for 72h.
Aalosa_ONT9