GSM2331847: H4; Homo sapiens; RNA-Seq
RNA was isolated from cells using the Trizol reagent (ThermoFisher), and purified according to the manufacturer’s protocol. All purified RNA was subsequently treated with DNaseI digestion to remove possible DNA contaminants (Qiagen). The quality of RNA used for cDNA library preparation was verified using the RNA nano 6000 analysis chip on a BioAnalyzer 2000 series instrument (Agilent Technologies) to ensure an RNA integrity value greater than or equal to 9. The cDNA libraries for sequencing were prepared from poly(A)+-selected mRNA, according to Illumina’s TruSeq Stranded mRNA sequencing preparation kit. Briefly, 1µg RNA was purified for mRNA. Then mRNA material was fragmented and denatured, in preparation for first and second strand cDNA synthesis steps. Finally, the 3’ ends were adenylated to ligate strand-specific adapter sequences to cDNA material and amplified using PCR.  Purity and size of cDNA library products were confirmed using a BioAnalyzer instrument. Library concentrations were determined via RT-qPCR using the Library Quantification Kit (KapaBiosystems). The samples were then prepared and sequenced on an Illumina HiSeq Series instrument, with one sample per sequencing lane to achieve > 2 x 108 reads per sample. stranded RNAseq
GEO Accession
GSM2331847