GSM5548016: iPSC_iMN1_sh_shTDP-43_rep1; Homo sapiens; RNA-Seq
Neuronal cultures were analyzed for RNA and protein 7 days post transduction. Total RNA was extracted 7 days post transduction using RNeasy Micro kit (Qiagen) per manufacturer's instructions, with lysate passed through a QIAshredder column (Qiagen) to maximize yield. RNA was quantified by Nanodrop (Thermo Scientific), with 75ng used for cDNA synthesis with SuperScript IV VILO Master Mix (Thermo Scientific). Splice variants in iPSC-derived motor neuron cells were established by PCR amplification from UNC13A exon 19 to exon 21 (UNC13A_19_21 FWD 5'-3'= CAACCTGGACAAGCGAACTG, UNC13A_19_21 RVS 5'-3'= GGGCTGTCTCATCGTAGTAAAC). Resulting products were purified using Wizard SV Gel and PCR Clean-Up columns (Promega) and submitted for Sanger and NGS (Amplicon EZ) (Genewiz).