LAO-A02 2010-10-04 Metatranscriptomic Sequence Data
Purified RNA was ethanol precipitated and stored at -80 oC overlayed with RNAlater solution (Ambion). Prior to sequencing library preparation, the pellet was rinsed twice in 80% ethanol and twice in 100% ethanol to remove any excess RNAlater solution. The pellet was then left on ice to dry. After ethanol evaporation, RNA pellets were resuspended in 1mM sodium citrate buffer at pH 6.4. Ribosomal RNAs were depleted using the Ribo-Zero Meta-Bacteria rRNA Removal Kit (Epicentre) according to the manufacturer's protocol. Transcriptome libraries were subsequently prepared using all synthesised cDNA with the ScriptSeq v2 RNA-Seq Library Preparation Kit (Epicentre) according to the manufacturer’s instructions.