GSM8013565: pbs3 [snRNA-seq]; Salmo salar; RNA-Seq
ScRNA-seq: Head kidney was sampled and placed in L15 (Leibovitz-15 + 5% FBS + 4 mM L-glutamine + 1% penicillin/streptomycin/amphotericin) and kept on ice. Head kidney tissue was passed through strainer (70 µm) in 10 mL PBS. Cell suspension filtered through a 30-µm strainer and centrifuged. Cells collected from the interface after Percoll gradient were washed and prepared in DPBS with BSA for subsequent sequencing. SnRNA-seq: Flash frozen head kidney tissue was minced in 1 mL TST buffer (2 mL of 2X ST buffer, 120 µL of 1% Tween, 20 µL of 2% BSA, 1.86 mL nuclease-free water). The homogenate was filtered through a 40-µm cell strainer and 3 mL of 1X ST solution (2X ST buffer [146 mM NaCl, 10 mM Tris-HCl pH 7.5, 1 mM CaCl2, 21 mM MgCl2, brought up to 10 mL with nuclease-free water]) was added. The samples were ready for sequencing after centrifugation, re-suspension in 1X ST buffer and filtration. Single Cell 3′ Reagent Kit v3.1 from 10X Genomics