title: Illumina HiSeq 4000 paired end sequencing; A universal protocol for isolating retinal ON bipolar cells across species via fluorescence-activated cell sorting - RNAseq of cell populations
design: A universal protocol for isolating retinal ON bipolar cells across species via fluorescence-activated cell sorting - RNAseq of cell populations
library_construction_protocol: NHP sample collection: Retinas were extracted from naïve NHP eyes and carefully dissected into quadrants to be cultured as explants. Each quadrant was independently processed for cell sorting and lead to the isolation of two cell populations, namely Goα+/Pcp2+high and Goα+/Pcp2+low. Mouse sample collection: Retinas were collected from naïve eyes of Rd1 wild type as well as Rd1 mutant mice. Following preparation for cell sorting, the two populations Goα+/Pcp2+high and Goα+/Pcp2+low were isolated.  Total RNA was extracted with an AllPrep® DNA/RNA FFPE Kit (#80234, Qiagen, Hilden, Germany) according to the manufacturer's instruction. Total RNA was submitted to the University of Manchester Genomic Technologies Core Facility (GTCF). Quality and integrity of the RNA samples were assessed using a 2200 TapeStation (Agilent Technologies). Illumina TruSeq Stranded mRNA kit, according to the manufacturer's protocol