RNA-seq of enriched dsRNA of T. vaginalis isolate G3
RNA-seq of enriched dsRNA of T. vaginalis G3, purchased from ATCC and minimally cultured; total RNA isolated using TRIzol, ssRNA depleted with 2M LiCl at -20C overnight, final dsRNA enrichment using DNase I and S1 nuclease to digest DNA and remaining ssRNA. Enriched dsRNA was shipped overnight on dry ice to Quick Biology (Pasadena, CA) for sequencing. Library for dsRNA-seq was prepared according to KAPA KK8540 RNA HyperPrep kit with 201-300 bp insert size (KAPA Biosystems, Wilmington, MA) using 25-50 ng total dsRNA as input. Final library quality and quantity were analyzed by Agilent Bioanalyzer 2100 and Life Technologies Qubit 3.0 Fluorometer. 150 bp PE (paired end) reads were sequenced on Illumina HiSeq 4000 (Illumina Inc., San Diego, CA). Sequences were demultiplexed to remove barcodes, and then adapters and other technical sequences were trimmed using TrimGalore v0.6.3 with the following parameters: paired, stringency=5, quality=20. PhiX positive-control spike-in reads were depleted using BWA-mem with default parameters.