GSM7117285: xh_220607_20-22h_mix; wastewater metagenome; RNA-Seq
Raw influx was filtered by 2 µm and 0.2 µm. Filtrate was either concentrated using Centricon units (60 ml), or used for Ceres nanotrap based extraction according to the manufacturer's protocol (10 ml) with addition of the ER2 reagent. RNA was extracted using Trizol/Trizol LS (Thermo Fisher) and the Zymo DirectZol kit (cat# R2050), or with the Zymo Biomics RNA Magbead kit (cat# R2137) using either the RNA isolation protocol from this kit's manual or the RNA/DNA combination isolation protocol from the manual of the Zymo Biomics MagBead DNA/RNA kit (cat# R2135). DNA was extracted using the Zymo RNA Magbead kit (cat# R2137) with the RNA/DNA combination protocol from the manual of the MagBead DNA/RNA kit (cat# R2135), or with the Zymo Biomics DNA Miniprep Kit (cat# D4300) RNA sequencing libraries were constructed using the SMARTer Stranded Total RNA-Seq Kit v3 - Pico Input Mammalian (Takara). DNA sequencing libraries were prepared by first applying the ResolveDNA Whole Genome Amplification kit (BioSkryb) and subsequently the Nextera XT DNA Library Preparation Kit (Illumina). A subset of RNA sequencing libraries was subjected to enrichment using the xHYB adventitious agent panel (Qiagen) where indicated in the sample description.