GSM8567180: RYm17, PBMC, aIL10_aPD1, wk0postATI; Macaca mulatta; RNA-Seq
Total RNA was isolated using the RNeasy Micro Kit (QIAGEN, Hilden, Germany) with on-column DNase digestion. mRNA purification was performed using magnetic beads with Oligo (dT). The mRNA was then mixed with a fragmentation buffer to generate short fragments. cDNA was synthesized using these mRNA fragments as templates. The short fragments were purified and resolved with EB buffer for end repair and single nucleotide A (adenine) addition. Adapters were ligated to the short fragments and suitable fragments were amplified via PCR. After quantification and qualification the sample libraries were pooled at equimolar concentrations and sequenced.