NextSeq 550 paired end sequencing; RNA-seq of brain organoids from human NKX2.1(GFP/w) hESCs treated with patterning molecules
RNA-seq of brain organoids from human NKX2.1(GFP/w) hESCs treated with patterning molecules
Individual organoids were flash frozen in isopentane on dry ice at day 14 and stored at -80. Samples were thawed in 'RNAlater-ICE (Thermo Fisher) at -20 overnight prior to RNA extraction. Patterning factors were added in low and high concentrations from day 3 to day 6 of organoid development. RNA extraction with Trizol and 1-Bromo-3-chloropropane, bead based RNA purification (Thermo Fisherâ€™s MagMax kit - spin procedure) Bulk RNA-seq libraries were constructed using the Smartseq2 protocol. Individual samples were indexed with Nextera XT Index kit v2, set B.