GSM5532183: PHH6-rep2-1B3nega; Homo sapiens; RNA-Seq
For selection of pericentral cell population cells were stained in PBS containing 1% BSA with SLCO1B3 antibody (NBP2-36492AF700) at 1:33 dilution for a minimum of 30 min. After the incubation, cells were washed three times and used for bulk sorting. 500,000 cells were sorted into15ml tubes for RNA isolation using Becton Dickinson Aria with a 130 µm nozzle. RNA samples were isolated using the same kit following the manufacturer's recommendations. 100-300 ng of RNA was used for cDNA synthesis using iScript cDNA Synthesis Kit (Bio-Rad, 1708891) following the online manual. 100 to 250 ng of total RNA was used for library construction using KAPA RNA Hyper + RiboErase HMR kit (Roche, 08098140702) following the manual. After the library construction, libraries were quantified on the bio-analyzer using DNA high-sensitivity kit (Agilent, 5067-4626), and sequenced paired-end (2 x 38bp) on the Illumina NextSeq 500 platform.