GSM6448514: KO52_dnFOS_RNAseq_rep2; Homo sapiens; RNA-Seq
Total RNA was extracted with NucleoSpin RNA (Machery-Nagel) and 1 ug of nucleic acid was retrotranscribed using SuperScriptTM II Reverse Transcriptase kit (Thermo Fisher) and 0.5 μg of Oligo(dT) primers (Promega), according to the manufacturer's instruction. RNA-seq libraries were prepared with a Total RNA Ribo-zero library preparation kit (with ribosomal RNA depletion) (Illumina, USA) according to manufacturer's instructions with the following alterations: 15 cycles of PCR was undertaken to amplify the library and adaptors for multiplexing were used at a 1:4 dilution. Library quality was checked by running the samples on a Bioanalyser and libraries were quantified using a Kapa library quantification kit (Kapa Biosystems, USA) and run on a NextSeq 500 with 75bp paired-end reads