GSM7623525: PBMC, BDBV challenged, VSV-BDBV-GP treated, Survivor 4, DPI 1, tech rep 1; Macaca fascicularis; RNA-Seq
At least three million PBMC were inactivated with Trizol LS buffer (Thermo Fisher Scientific) for isolation of RNA. RNA was isolated using Direct-zol RNA Miniprep purification kits (Zymo Research) following the directions of the manufacturer with the optional DNAse I treatment and subsequent wash steps. RNA was eluted in 30ul of DNase/RNase-free water. An Illumina Ribo-Zero Stranded kit was used to deplete ribosomal RNA (rRNA) and construct cDNA libraries according to the protocol provided by the manufacturer. RNA was fragmented, converted to double-stranded cDNA, and adapters ligated to each strand. The resulting base-pair cDNA fragments were then amplified by PCR. Each library was prepared with a unique indexed adapter for multiplexing. Multiplexed libraries were subjected to paired-end 75 base pair sequencing using the Illumina NextSeq550 PE platform.