Illumina 16S metabarcoding of DNA-5BS
In order to obtain the co-isolation of RNA and DNA from the same soil sample, the PowerSoil total RNA kit (Qiagen) and PowerSoil DNA elution kit (Qiagen) were used following the manufacturer's recommendations. Briefly, three grams of soil were used as an initial sample, and the mechanical lysis was performed with a mixture of glass beads and bead solution processed by three pulses of 30 seconds at 6 m/s in a FastPrep-24 (MP Biomedicals). Nucleic acids were quantified by fluorometry using a specific Qubit dsDNA BR Assay Kit (Invitrogen) for DNA samples and Qubit RNA BR Assay Kit for RNA samples. Integrity of the DNA samples was corroborated in a 1% agarose gel, while an agarose/formaldehyde gel was used for the RNA samples. The DNA and RNA samples were stored at -20C and -80C, respectively, until use. The V1-V3 region of the bacterial 16S rRNA gene was amplified with the primers 28F (5-GAGTTTGATCNTGGCTCAG-3) and 519R (5-GTNTTACNGCGGCKGCTG-3). The amplicons were sequenced with an Illumina sequencer using the service of Molecular Research LP. (Mr. DNA Lab). In the case of the RNA samples, reverse transcription was performed prior to amplification to generate cDNA.