Metagenomics of Hymeniacidon heliophila host microorganisms
The primer pair 515F-806R was used to amplify the V4-region of the 16S rRNA gene of Bacteria and Archaea using the extracted DNA (Apprill et al., 2015; Parada et al., 2016) and aliquots were sequenced at the NGS Solues Genmicas (Piracicaba, SP. Brazil). The reaction mixture (25 L) was prepared with 2.5 L of template DNA (20 ng/L), 0.20 nM of both primers and 2X PCRBio Ultra Mix (PCRBiosystems). The thermal cycle was set as following: 3 min at 95C, 30 cycles of 30 s at 95C, 30 s at 60C and 30 s at 72C. Final extension was performed for 10 min at 72C (Hardoim et al., 2021). The amplicons obtained were subjected to Illumina sequencing using MiSeq platform