metatranscriptomics of Homo sapiens: adult colonic mucosa
RNA was extracted from human colonic mucosa tissues using the ZymoBIOMICS DNA/RNA Miniprep Kit (Zymo Research, Irvine, CA) and was library prepped using the NEB Next Ultra II RNA Library Kit for Illumina (New England BioLabs, INC, Ipswhich, MA). Equimolar ratios of each library were pooled and gel-purified using 2% agarose gels and GelSTAR Nucleic Acid Stain (Lonza Bioscience, Walkersville, MD) using the QIAquick (QIAGEN, Germantown, MD) gel purification kit. The final purified library quality was assessed using the High Sensitivity DNA Assay on a Bioanalyzer 2100 (Agilent, Santa Clara, CA) before being sequenced in two runs; first on an Illumina HiSeq 4000, generating PE150 reads (UC Davis, CA), and samples with low sequence counts were rerun on an Illumina NovaSeq 6000, using a SP300 flow cell to generate PE150 reads (UC Davis, CA).