title: Metagenomics (MiSeq-Illumina) biomass anaerobic reactors in the surfactant degradation (LAS)
design: The gene?specific sequences used in this protocol target the 16S V3 and V4 region. They are selected from the Klindworth et al. publication (Klindworth A, Pruesse E, Schweer T, Peplles J, Quast C, et al. (2013) Evaluation of general 16S ribosomal RNA gene PCR primers for classical and next?generation sequencing?based diversity studies. Nucleic Acids Res 41(1).) as the most promising bacterial primer pair. Illumina adapter overhang nucleotide sequences are added to the gene?specific sequences. The full length primer sequences, using standard IUPAC nucleotide nomenclature, to follow the protocol targeting this region are: 16S Amplicon PCR Forward Primer =5' TCGTCGGCAGCGTCAGATGTGTATAAGAGACAGCCTACGGGNGGCWGCAG 16S Amplicon PCR Reverse Primer =5' GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAGGACTACHVGGGTATCTAATCC (in red is the sequence of the adapters). The sequencing was done in Miseq - Illumina, 2x250 cycles