title: RNA-seq (Metatranscriptomics) of Jejunal contents of Gallus gallus
design: Total RNA was extracted from jejunal content samples using the RNeasy PowerFecal Pro Kit (Qiagen, Hilden, Germany). RNA quality was assessed using the High Sensitivity RNA Tapestation (Agilent Technologies Inc., California, USA) and concentration was determined using the AccuBlue Broad Range RNA Quantitation assay (Biotium, California, USA). Ribosomal RNA was depleted using QIAseq FastSelect HMR+5S/16S/23S (Qiagen, Hilden, Germany) per manufacturers instructions to enrich for total mRNA. The library was prepared according to the NEBNext Ultra II Directional RNA Library Prep Kit (New England BioLabs Inc., Massachusetts, USA) with Illumina 8-nt dual-indices generating libraries of approximately 300 bp. Library concentration was measured by a Qubit 2.0 fluorometer (ThermoFisher, Massachusetts, USA) and quality was assessed using the TapeStation D1000 ScreenTape assay (Agilent Technologies Inc., California, USA). Equimolar pooling of libraries was performed based on quality control values, and sequencing was performed on a DNBSEQ T7 platform to produce 40M paired-end of 150bp-reads per samples.
identifiers.namespace: SUB15758866
identifiers.id: JC223D