title: Dengue virus sequencing after infecting Wolbachia-contained C6/36 cells
design: , viral genomes in the extracellular particles were also analyzed. Supernatants (~10 ml) from cell cultures in T-75 flasks were centrifuged at 15,000 g for 45 minutes in 40 % PEG8000 solution containing 10 mM Tris (pH 8.0), 120 mM NaCl and 1 mM EDTA (68). Viral particles in the pellets and the remaining 0.5 ml solutions were thoroughly mixed with 1.5 ml Trizol-LS (Ambion) and 0.4 ml chloroform. After the centrifugation at 12,000 g for 15 minutes, the upper layer separated from Trizol/chloroform (bottom layer) was used for ethanol precipitation to collect viral RNA. The pellet was washed with 70% ethanol, dried, and dissolved by Tris (10 mM, pH8.0)-buffered DEPC-water. The RNA was selected with the RNA Clean & ConcentratorTM kit (Zymo Research), containing DNase. Library preparation and RNA-seq sequencing were performed at Zymo Research Inc. (Irvine, CA),
identifiers.namespace: SUB15202903
identifiers.id: DENV1_5days_C6/36 cells