limonene degradation
Genomic DNA was amplified with primers specific to the V3-V4 region of the 16S rRNA gene which also incorporates the Illumina overhang adaptor. Paired-end type reads with 500 bp fragments were generated and amplified using the primers pair 341F and 806R (Fouhy et al., 2015). After quantification by the Real Time PCR methodology using Kit KAPA-KK4824 (Library Quantification Kit - Illumina / Universal), an equimolar pool of DNA was generated, by normalizing all samples at 2nM, to perform the sequencing, which was conducted using the new generation Illumina MiSeq sequencing system (Illumina Sequencing). The methodologies were compatible with the manufacturer's protocol. For the bioinformatics study, the GREENGENES database was used, using the QIIME software (CAPORASO, et al. 2010).
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