GSM7925065: Rhesus_2 [SPB1EB1A]; Macaca; RNA-Seq
After incubation, blood in TruCulture tubes was separated according to the manufacturer's instructions with additional centrifugation at 800 ×g for 10 minutes.  Supernatants were saved and frozen at −80 °C for cytokine analysis.  The cell pellets were lysed using red blood cell lysis solution for 10 min followed by centrifugation at 2,000 ×g for 3 minutes, and remaining white blood cells were lysed in Qiazol and frozen at −80 °C until library construction (Qiagen). RNA isolated from white blood cells (Qiazol, Qiagen) was quality controlled using A260/A280 ratio, 28S/18S rRNA ratio, and an RNA integrity summary score (Agilent RIN). Total RNA samples (250ngs, RIN>=7.0) were enriched for mRNA, fragmented, and converted into indexed cDNA libraries according to Illumina TruSeq Stranded mRNA protocol and quantity controlled using Agilent TapeStation. Samples were sequenced to at least 25M of 2 x 50bp paired-end reads and assessed using in-house QA/QC metric (Q2Solutions Expression Analysis).