GSM9512031: Zebrafish,hsd17b7_ctrl; Danio rerio; RNA-Seq
Inner ear and lateral line sensory hair cells were isolated from 4 days post-fertilization (dpf) Tg(Brn3c:mGFP) zebrafish larvae of both wild-type and hsd17b7 mutant backgrounds. Tissues were enzymatically dissociated using trypsin to generate single-cell suspensions, followed by centrifugation, resuspension, and cell counting. GFP-positive cells were enriched by fluorescence-activated cell sorting (FACS) prior to single-cell RNA sequencing library preparation. Single-cell RNA sequencing libraries were prepared using the 10x Genomics Chromium Single Cell 3′ v4 reagent kit according to the manufacturer's instructions. Briefly, single-cell suspensions were loaded onto Chromium microfluidic chips, where individual cells were encapsulated with barcoded gel beads in oil emulsions (GEMs) using a Chromium Controller (10x Genomics). Barcoded RNA was reverse-transcribed, and sequencing libraries were constructed following standard 10x Genomics protocols.