GSM5582226: unaffected 1; Homo sapiens; RNA-Seq
RNA extraction using QIAamp RNA Blood Kit (Qiagen) Illumina TruSeq stranded total RNA with ribo-zero globin Ten micrograms of total RNA was used to generate index-tagged paired-end cDNA libraries. Sample preparation was performed using TruSeq stranded total RNA with ribo-zero globin sample preparation kit (Illumina). Briefly, rRNA and globin RNA was depleted using Globin Removal Mix (Illumina). RNA fragmentation was performed using RNA Fragmentation Reagents (Life Technologies) to obtain 200–300 bp fragments. First strand cDNA was generated using the Superscript II Reverse Transcriptase and random hexamers (Life technologies). Second strand cDNA was synthesized and 3' ends adenylation was performed (Illumina). Illumina sequencing adapters were ligated to cDNA and PCR amplification was performed on the adapter-ligated cDNA with 2X Phusion DNA polymerase Master Mix (New England Biolabs). Samples were sequenced 100bp (2×100) paired-end on Illumina HiSeq2500