Thermophilic_T3_rep1_Microbial community AD_Metagenomics
Digestate samples were collected at T0 (initial inoculum), T3 (end of the wash out phase), T5 (middle of the stable phase) and T6 (end of the stable phase) for all reactors. All samples were pelleted at 13,000 rpm for 20 min. From 250 mg of each pellet, DNA was extracted using the DNeasy PowerSoil Kit (Qiagen, Germany) according to the manufacturers instructions. DNA yields were quantified using a Qubit 1dsDNA Broad Range assay kit (Thermo Fisher Scientific), purity was assessed on a Nanodrop 1000 spectrophotometer (Thermo Fisher Scientific), while fragmentstion was determined through gel electrophoresis 1 % (w/v) 1  TAE agarose gels. DNA was stored at 80 C until analysis. Shotgun Metagenomics (10G raw data per sample NovaSeq X Plus Series (PE150)) was performed at BMR Genomics (Padova, Italy).