RNA-Seq for homo sapiens: serum
Total RNA was purified from 200 L of serum, using PureLink Viral RNA/DNA Mini Kit (Invitrogen, USA), following manufacturers recommendations. cDNA was synthesized using random primers and the SuperScript IV Reverse Transcriptase (Thermo, USA), following the manufacturers recommendations. The cDNAs were mechanically fragmented, quantified with the Qubit 2.0 (Life Technologies, Carlsbad, USA) and analyzed with the Bioanalyzer 2100 (Aligent, Santa Clara, USA). Sequencing libraries were prepared using the Nextera XT DNA Library Preparation Kit (Illumina, San Diego, USA), according to the manufacturers recommendations. The samples were labeled with adapter sequences using Nextera XT Index Kit (Illumina, San Diego, USA), according to the manufacturers recommendations. Paired-end sequencing was performed using the HiSeq 2000 Sequencing System (Illumina, USA) and the MiSeq Reagent Kits v2 (600 cycles) (Illumina, USA).