16S rRNA SEQUENCING Brachionus plicatilis CULTURE: DAY 7
The water RNA/DNA Purification Kit (Norgen Biotek Corp.) was used for extracting total DNA from the filters, following the manufacturer's specifications. DNA quality, integrity, and concentration were verified through electrophoresis on a 1% agarose gel, NanoDrop 2000 (Thermo Fisher Scientific), Qubit4 Qubit dsDNA BR Assay Kit (ThermoFisher Scientific), and TapeStation 4150 capillary gel electrophoresis system (Agilent Technologies). The V3-V4 region from the 16S rRNA gene was PCR amplified using the primers Bakt-341F (5-CCTACGGGNGGCWGCAG-3) y Bakt-805R (5-GACTACHVGGGTATCTAATCC-3). The kit Herculase II Fusion DNA Polymerase Nextera XT Index V2 was used to generate the amplicon library. The library size distribution was screened with Agilent 2100 TapeStation system and D1000 ScreenTape, and quantified using qPCR according to the Illumina qPCR quantification protocol guide. Sequencing was done with the Illumina MiSeq system 2 x 300 bp  (Macrogen Inc.)