GSM9319456: 5m_S1; Aedes albopictus; RNA-Seq
Total RNA was extracted from wAlbB-infected C6/36 cells using the miRNeasy Mini Kit (Qiagen, Hilden, Germany), with protocol adaptations. Samples were incubated at 37 °C in a heating block until completely thawed, ensuring that all salts were fully dissolved. After an additional 5 min incubation at room temperature, samples were mixed with 100 µl 1-Bromo-3-chloro-2-propanol (BCP, Tokyo Chemical Industry, Tokyo, Japan). The sample-BCP mixture was vigorously shaken for 15 seconds, followed by a 2–3 min incubation at room temperature. Phase separation was achieved through a 15 min centrifugation at 12,000 g, 4 °C. The upper aqueous phase, containing the nucleic acids, was retained, with 350 µl applied to the Qiagen column. The extract was also subject to DNase I treatment to eliminate contaminating DNA. Pilot 1: Three samples were subjected to rRNA depletion using Illumina TruSeq Stranded Total RNA with Ribo-Zero Plus rRNA Depletion according to the manufacturer's instructions, using the maximum input of 5 µg RNA. Pilot 2: Four libraries corresponding to two technical replicates.