16S rRNA amplicons from chlorotoluron DNA-SIP of biofilter metagenome
To generate 16S rRNA gene amplicons for sequencing, a nested PCR was performed involving the product of a first PCR with primer set 27F/1492R (Turner et al., 1999). The obtained amplicons were purified using the QIAquick PCR Purification Kit (Qiagen, Germany) and used as template for the second PCR. The latter targeted either the V4 region of the 16S rRNA gene using primers A519F and 802R (Klindworth et al., 2013) or V3-V4 region using primers Bakt_341F and Bakt_805R (Klindworth et al., 2013) amended with overhang adapter sequences. The amplicons were obtained and purified according to the Illumina 16S Metagenomic Sequencing Library Preparation protocol (Illumina, San Diego, CA).
SUB9311058
12C_OFO_CLT_t1_heavy_2