Single-cell RNA-seq of Danio rerio: Transgenic FLT3-ITD IDH2-R172K
Single cell RNA-seq library.Single viable KM cells were collected from the transgenic double mutant (n=3; pooled) and WT (n=3; pooled) fish at 7 months old into 0.9X PBS with 5% FBS. Their viability was examined by 0.4% Trypan blue staining under microscopy. The single-cell library was constructed using the ChromiumTM Controller and ChromiumTM Next GEM Single Cell 3 Kit v3.1 (10x Genomics, Pleasanton, CA). Complementary DNA (cDNA) was synthesized from the fragmentated RNAs using N6 random primers, followed by end repair and ligation to BGISEQ sequencer compatible adapters. Quality control of the final library was performed by checking the distribution of the fragments size using the Agilent 2100 bioanalyzer and quantification was performed by real-time quantitative PCR using TaqMan probes. The final products were sequenced using the DNBSEQTM platform (BGI- HK, China).