title: GSM4628143: scRNA-seq zebrafish 4dpf, neuromast hair cells; Danio rerio; RNA-Seq
library_construction_protocol: Zebrafish larvae Tg(myo6b: RiboTag) at 4 dpf were dissected to remove the head, which included the ear. Approximately 400 dissected bodies in batches of 50 were dissociated by adding 0.5 ml of 0.5% trypsin (Gibco, cat# 15400054) and triturated with 1 ml pipette tip for 30 seconds at 30°C. The dissociated cells were filtered with 40 μm strainer (Pluriselect, cat# 43-50040-50) and washed with PBS twice before adding DMEM containing 10% FBS and spun down by centrifugation at 700g rpm for 5 min at 4°C. Cells were resuspended and kept in DMEM before processing for FACS. scRNA-seq was carried out with 10X Genomics single cell platform (10X Genomics, Pleasanton, CA. USA). Approximately 5000 live cells after FACS in a maximum volume of 34 microliters were loaded on a 10X Genomics Chromium Controller (10x Genomics). Chromium Single Cell 3' Library and Gel Bead Kit v2 (10X Genomics) was used for libraries preparation according to manufacturer's instructions. Quality of the libraries was evaluated on a Fragment Analyzer instrument (Agilent) and sequenced on a NextSeq 500 sequencer Illumina with the following paired read lengths: 26 bp Read 1, 8 bp I7 Index and 98 bp Read 2.
attributes.tag: GEO Accession
attributes.value: GSM4628143