qPCR raw datasets on the assessments of cover crop monocultures and mixtures improving the rhizosphere bacterial abundance and functionality through rerooting

Quantitative PCR (qPCR) raw data includes the source data that corresponds to the the counts of 16S rRNA gene copies per gram of soil for different variations, as discussed in the research article - "Cover crop monocultures and mixtures improve the rhizosphere bacterial abundance and functionality through rerooting". The copy number of the 16S rRNA gene per gram of soil was quantified by SYBR® Green-based qPCR using a 7500 Fast Real-Time PCR System (Applied Biosystems™, Thermo Fisher Scientific, Waltham, MA, USA). Aliquots of the same DNA extract utilized in amplicon sequencing were used in qPCR. Dilutions of template DNA were used to compensate for the effect of PCR inhibitors in the samples. Each sample was analyzed in triplicate. A PCR amplicon of the Escherichia coli V3 region was used as standard. Each reaction of 20 µL contained 1 µL of template DNA, the forward primer 341F (Muyzer et al., 1993), the reverse primer 518R (Muyzer et al., 1993), and Luna® Universal qPCR Master Mix (NEB). Reaction conditions were an initial denaturation for 1 min at 95 °C, followed by 40 cycles of denaturation at 95 °C for 15 s and extension at 60 °C for 30 s. The melting curve was recorded in the temperature range of 60 °C to 95 °C. The 16S rRNA gene copy numbers per gram of soil were calculated using the standard curve method and then normalized against the standard (Adelowo et al., 2018). The average efficiency value was 100.77 ± 3.15 %. The absolute copy numbers for each bacterial phylum were calculated by multiplying the qPCR values by the relative abundance values in percent obtained from the 16S rRNA gene sequencing analyses.