RNA-seq of ALS patients
Total RNA was isolated from fibroblasts using RNeasy Mini Kit (Qiagen). RNA integrity was assessed using RNA BR Assay Kit on Qubit 2.0 and RNA 6000 Pico Kit on Bioanalyzer 2100 (Agilent). 500 ng of RNA at integrity number > 8 was converted to cDNA using TruSeq Stranded Total RNA with Ribo-Zero kit (Illumina). Libraries were assessed qualitatively on Bioanalyzer 2100 using High Sensitivity DNA Kit and quantitatively on CFX96 Real-Time PCR system (Bio-Rad) using KAPA Library Quantification Kit (Kapa Biosystems), and then sequenced 2x150 bp on HiSeq2500 Illumina platform.