GSM3415975: HJ4_ML02-1_S151; Homo sapiens; RNA-Seq
Cells were lysed in Trizol and stored at -80C until RNA isolation. Total RNA was extracted using a Direct-zol™ RNA Kit (Zymo Research, cat#:R2051) according to the manufacturer's instructions. The extracted RNAs were quantified and quality checked using a NanpDrop 1000 Spectrophotometer (Thermo Fisher Scientific) and a BioAnalyzer RNA 6000 Nano Kit (Agilent Technologies). The cDNA libraries were prepared using the NEXTflex™ Rapid Directional RNA Sequencing Kit and NEXTflex™ RNA-Seq Barcodes – 24 Kit (BioO Scientific) as per the manufacturer's instructions. The final product was assessed for its size distribution using Bioanalyzer DNA High Sensitivity Kit (Agilent Technologies) and for its concentration using Kapa library quantification kit (Kapa Biosystems). Libraries were pooled equimorlarily, followed by on-board cluster generation on a Rapid Run single-end flow cell and subsequent 50 cycles sequencing (v3 sequencing kit) according to the manufacturer's instructions (HiSeq 2500, Illumina).