title: 16S rRNA gene amplicons of the DNA-SIP samples obtained from a biofilter microcosm prior to fractionation; timepoint 2
design: To generate 16S rRNA gene amplicons for sequencing, a nested PCR was performed. The first PCR targeted the V1-V5 region of the bacterial 16S rRNA gene using primers GM3F and 908R (Klindworth et al., 2013). The obtained V1-V5 amplicons were purified using the QIAquick PCR Purification Kit (Qiagen, Germany) and used as template for the second PCR. The latter targeted the V4 region using primers A519F and 802R(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).
library_name: SIP2-PC