GSM9896498: HUB-FT-s003; Danio rerio; RNA-Seq
Unfixed ventricles were dissociated by Collagenase and TrypLE Express treatment to prepare single cell solutions. Cells were subsequently FACS sorted based on myl7:PercevalHR expression in 384 well plates, each well containing 1 cell, containing Vapor-Lock oil with CEL-Seq2 barcoded primers, spike-in mRNA and dNTPs (Single Cell Discoveries). Plates were spun down and frozen at -80oC until ready for library prep. Single-cell sequencing libraries were prepared by Single Cell Discoveries (Utrecht, The Netherlands) using the SORT-seq protocol. The CEL-Seq2 protocol was used for library prep (Hashimshony et al., 2016)). Illumina sequencing libraries were prepared with the TruSeq small RNA primers (Illumina) and paired-end sequenced at 75 bp read length on the NovaSeq X Plus Illumina sequencer. In total six 384-well plates were sequenced containing one cell per well of which three were obtained from wildtype cells and three from plna R14del mutant cells.