L5 metatranscriptomics reads
The L5 sample was frozen in liquid nitrogen and pulverized with an oscillating ball mill (TE-350, Tecnal Inc.). The homogenized sample was used for RNA extraction using the RNA PowerSoil Total RNA Isolation Kit (CAT: 12866-25 MoBio), according to manufacture instructions. The quality of RNA was verified using an Agilent Bioanalyzer 2100 with the RNA 6000 Nano Reagents Kit (Agilent). The RNA was treated with a blend of the Ribo-Zero rRNA removal Kit for bacteria and the Ribo-Zero Magnetic Gold Kit (Epicentre Biotechnologies) to remove both prokaryotic and eukaryotic rRNAs, respectively. Subsequently, the supernatant was purified using an 80% (v/v) ethanol solution, and the resultant RNA was used for library RNA preparation using the Truseq Total RNA kit (Illumina).