Illumina HiSeq 4000 paired end sequencing; RNA-seq of peripheral blood mononuclear cell (PBMC) derived from normal healthy donors and systemic lupus erythematosus (SLE) patients
RNA-seq of peripheral blood mononuclear cell (PBMC) derived from normal healthy donors and systemic lupus erythematosus (SLE) patients
PBMCs were isolated from fresh whole blood using Ficoll (Ficoll-Paque Plus, GE Healthcre). Samples of four (total eight) donors were pooled and Samples of four (total eight) SLE patients were pooled. Samples were washed twice with PBS and 1 mL of TRIzol per 1 x 107 cell pellets was added and frozen in -80℃. Blood was collected in heparinized tubes and a portion was removed for PBMC analysis.  Total RNA was extracted from 107 PBMCs using TRIzol reagent (Life Technologies).Total RNA from cells was purified using the RNeasy Mini Kit (Qiagen, Venlo, Netherlands). All RNA content in the samples was measured with a NanoDrop®ND-1000 spectrophotometer, and the RNA quality was assessed using RNA 6000 NanoChips with the Agilent 2100 Bioanalyzer (Agilent, Palo Alto, CA, USA). Libraries were prepared using the TruSeq Stranded Total RNA Library Prep Kit with Ribo-Zero Gold (Illumina).