title: GSM6260215: iPSC-neurons, 5 µM PTBP2 gapmer, rep3; Homo sapiens; RNA-Seq
library_construction_protocol: iPSC-derived neurons were lysed with TRIzol Reagent (Invitrogen 15596-026). Chloroform was added to the lysate, and samples were centrifuged at 4°C to separate phases. After addition of ethanol, the aqueous phase was applied to an RNeasy Mini column (Qiagen #74104). On-column DNAse treatment was performed using Qiagen DNAse Set (Qiagen #79254) at room temperature for 15 minutes. Purified total RNA was analyzed on Agilent TapeStation systems, using D1000 Screen Tape & Reagents, and then quantified with a Qubit BR kit (Thermo Fisher #Q10210). Poly(A)+ RNA transcript was isolated from 1 ug purified total RNA (RNA integrity number >9.0) with NEBNext poly(A) mRNA magnetic isolation module (New England Biolabs #E7490). RNA-seq libraries were prepared with NEBNext Ultra Directional RNA library preparation kit for Illumina (New England Biolabs #E7420S) according to the manufacturer's instruction.