Avocado waste anaerobic digestion microbiome
The total environmental DNA was extracted using Faecal/Soil Total DNA extraction kit (Zymo Research Corporation, CA, USA) according to manufacturers instructions. The quantity and quality of extracted DNA was checked on Biodrop Lite spectrophotometer (Biochrom Ltd, Cambridge, UK) and agarose gel electrophoresis, respectively. Extracted DNA used for downstream PCR and sequencing analysis had A260:A280 ratios between 1.82.0 and DNA concentrations of 20150 ng/L. Resultant DNA was employed with 16S rRNA gene fragments to execute PCR amplification using Quick-16S Primer Set V3-V4 (341F; 5-CCTACGGGNGGCWGCAG-3 and 785R; 5-GACTACHVGGGTATCTAATCC-3) (Zymo Research Corporation, CA, USA). Cleaning of the resultant PCR products, index library preparation, pooling and sequencing on Illumina Miseq 250 to generate paired 300-bp high-quality reads were performed at Inqaba Biotechnology (Pretoria, South Africa).