16s mbs1
A sample of surface soil was collected in the coastal area of Lázaro Cárdenas, Michoacán in March 2022. Environmental DNA was extracted using the DNeasyPowerSoil Pro Kit (QIAGEN) following the manufacturer's instructions. The V3-V4 region of bacterial 16S rRNA genes was amplified and sequenced using an Illumina MiSeq platform with a 300-bp paired-end format (Macrogen Co., Seoul, South Korea). The sequencing library is prepared by random fragmentation of the DNA sample, followed by 5' and 3' adapter ligation. Alternatively, "tagmentation" combines the fragmentation and ligation reactions into a single step that greatly increases the efficiency of the library preparation process. Adapter-ligated fragments are then PCR amplified and gel purified. For cluster generation, the library is loaded into a flow cell where fragments are captured on a lawn of surface-bound oligos complementary to the library adapters. Each fragment is then amplified into distinct, clonal clusters through bridge amplification. When cluster generation is complete, the templates are ready for sequencing.