Total RNA Sequencing of freshwater metagenomes: mesocosm experiment
DNA and total RNA were separately extracted from samples in 96-well plates using the NucleoMag DNA/RNA Water kit (D-MARK Biosciences, Toronto, Canada). DNA and RNA libraries of all samples and negative extraction controls were prepared for metagenomics and totalRNA-Seq using the NEBNext Ultra II DNA Library Prep Kit for Illumina and the NEBNext Ultra II Directional RNA Library Prep Kit for Illumina, respectively (New England Biolabs, Whitby, Canada). DNA and RNA library pools were paired-end (2x100 bp) sequenced in a 50:50 ratio on a single lane of a NovaSeq 6000 SP flowcell.