GSM4727528: Cerebral Organoids KO/WWOX 2; Homo sapiens; RNA-Seq
Total RNA was extracted from homogenized organoids using Trizol reagent and standard phenol-chloroform isolation technique. RNA quality was assessed by using RNA ScreenTape kit, D1000 ScreenTape kit, Qubit(r) RNA HS Assay kit and Qubit(r) DNA HS Assay kit. For mRNA library preparation, 1ug of RNA per sample was processed using KAPA Stranded mRNA-Seq Kit with mRNA Capture Beads. Library was eluted in 20µl of elution buffer, adjusted to 10mM, then 10µl (50%) from each sample was collected and pooled in one tube. Multiplex samples Pool (1.5pM including PhiX 1.5%) was loaded in NextSeq 500/550 High Output v2 kit (75 cycles) cartridge and loaded on NextSeq 500 System machine, with 75 cycles and single-read sequencing conditions.