GSM9087548: Library_1_liver_all_samples; Salmo salar; RNA-Seq
Samples were placed in a six well plate containing tween-salt-tris buffer (TST) and RNase inhibitor (Merck) and the nuclei dissociated by mincing the tissue for 10 minutes with spring scissors (Noyes) alongside intermittent pipetting. The solution was filtered through a 40µm Falcon Cell Strainer before nuclei centrifugation at 4°C for 5 min at 500g and resuspension in salt-tris solution. Nuclei were quality checked and fixed as per the manufacturer's instructions (Parse fixation kit 2.0) then counted using a haemocytometer (Neubauer DHC-N01). snRNA-seq libraries were generated from the fixed nuclei using a Parse Biosciences Evercode WT v2 Mini kit following the manufacturer's protocol. The final snRNA-seq libraries were equimolar pooled and sequenced on a NextSeq P3 (200 cycle) Illumina flow cell using the Parse Biosciences Illumina Run Configuration (Read 1: 74 cycles, i7 Index: (Index 1) 6 cycles, Read 2 (barcode): 86 cycles, i5 Index (index 2): 0 cycles) by the University of Edinburgh's Genetics Core Facility (Edinburgh Clinical Research Facility).