GSM7567185: Chronic infection with CNS pathology (CP+)-BH-1; Macaca nemestrina; miRNA-Seq
bdEVs were separated from frozen occipital cortex tissues using our published protocol with minor modifications (Huang et al., 2022a, 2022b, 2020). Before extraction, a small (∼50 mg) piece of tissue was stored at –80°C for later RNA extraction from brain homogenate (BH).  bdEV RNAs were extracted by adding Trizol LS (Thermo Fisher 10296028) to 100 μL bdEV resuspension. After phase separation, RNAs were purified by miRNeasy Mini Kit solutions (Qiagen217004) and Zymo-Spin I Columns (Zymo ResearchC1003–50) according to the manufacturer's instructions.  BH RNA was extracted by adding Trizol (Thermo Fisher15596018) and homogenizing tissues with Lysing Matrix D (MP Biomedicals116913100) in a benchtop homogenizer (FastPrep-24, MP Biomedicals) at 4.0 m/s for 20 seconds. After homogenization, the supernatant was collected, and RNA was isolated by miRNeasy Mini Kit solutions (Qiagen217004) and Zymo-Spin IIICG Columns (Zymo Research C1006-50-G) following the manufacturer's instructions. RNA was resuspended in 40 μL Rnase-free water, and 8 µl was used for small RNA libraries construction by the D-Plex Small RNA-seq Kit (Diagenode C05030001). Indexes were attached using the D-Plex Single Indexes for Illumina - Set A (Diagenode C05030010) according to the manufacturer's protocol.