GSM5764629: ALS_Affected_3_Week_Replicate_3; Homo sapiens; RNA-Seq
Frozen cell pellets were resuspended in Qiagen's RLT Plus +beta-mercaptoethanol solution and extracted according to the All Prep DNA/RNA Mini Kit (Qiagen) RNA was quantifed by fluorimetric methods (Qubit, Invitrogen) and quality assessed by Fragment Analysis (Agilent).  Library preparation was initiated with 500ng of RNA and completed following manufacturer's recommendations for the TruSeq stranded mRNA kit (Illumina).  Final libraries were quantified by qPCR (Kapa Library Quantification Kit, Kapa Biosystems) and fragment size observed with high sensitivity DNA fragment analysis (Agilent).  Paired end sequencing was completed on the NovaSeq 6000 at 2x100bp reads, targeting >50 million reads per sample.