16S bacteria amplicon seq of droplet enrichment of groundwater PNK149 with Anammox necromass and soil extract, day 0, replicate 1
PCR amplification of the bacterial V3-V4 region of the 16S rRNA was performed in a 25 L reaction. Illumina sequencing adapters TCGTCGGCAGCGTCAGATGTGTATAAGAGACAG and GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAG were attached at the 5 ends of the forward and reverse primers, respectively. PCR was conducted using primer combinations Bakt_0341F/Bakt_0785R (Klindworth et al., Nucleic Acids Res. 2013;41(1):111.). The cycling conditions for 16S rRNA with an initial denaturation at 98C for 3 min, 40 cycles of denaturation at 98C for 30 s, annealing at 55C for 40s, elongation at 72C for 1 min, and final extension at 72C for 5 min. The amplicons were sequenced on the Illumina MiSeq platform with 2 x 300 bp paired-end by IIT Biotech (Bielefeld, Germany).