Microorganismos en digestatos de plantas de biogas

Abstract

Microbiological analysis Samples for microbiological analysis were collected prior to the fraction separation step from the three plants under study. They were stored at -20 °C before DNA extraction. DNA was extracted from each sample using the FastSpin for Soil Kit (MP Biomedicals, USA) according to the manufacturer's protocol. Sequencing was performed at the Genomics Unit of the Biotechnology Institute of INTA (National Institute of Agricultural Technology). Sequencing of 16S rRNA in hypervariable region V3-V4 was performed with universal primers 341F (CCTACGGGRSGCAGCAGCAG) and 805R (GACTACHACHVGGGGGTATCTAATCC). The sequencing method was 2x300 PE-reads on Illumina MiSeq. Data were analyzed using the DADA2 package in R to form ASVs (Amplicon Sequence Variants) (Engelbrektson et al., 2010; Bolger et al., 2014).