GSM3463814: 28501; Homo sapiens; RNA-Seq
RNA used for RNA sequencing was purified using a miRNeasy mini kit (Qiagen, CA, USA) and the RNA used for PCR analysis using RNeasy mini kit (Qiagen, CA, USA) according to manufacturer's protocol with on-column DNA digestion (Qiagen). The concentration and quality of the RNA was assessed with a NanoDrop ND-1000 spectrophotometer (NanoDrop Technologies, Wilmington, DE, USA). Agilent Bioanalyzer RNA nano chips (Agilent) were used to evaluate the integrity of total RNA and Qubit RNA –kit (Life Technologies) to quantitate RNA in samples. 1 µg of total RNA was used for ScriptSeq™ Complete Gold System (Epicentre) to ribodeplete rRNA and further for RNA-seq library preparation. SPRI beads (Agencourt AMPure XP, Beckman Coulter) were used for purification of RNAseq libraries. The library QC was evaluated on High Sensitivity chips by Agilent Bioanalyzer (Agilent).