title: 16S amplicon seq of soil samples from microcosms without nematodes, incubated with 16O-water for 5 days, replicate 2, DNA-SIP density fraction 8
design: Polymerase chain reaction was performed using primer pair Bact_341F/Bact_805R (Herlemann et al., 2011) and Ruby Taq Mastermix (Jena Bioscience GmbH, Jena, Germany). The primers were modified with a 5'-overhang as adaptor for Illumina sequencing. Amplicons were purified using NucleoSpin Gel & PCR Clean-Up Kit (Macherey-Nagel, Dren, Germany).  Barcodes were then added in a second PCR according to the Illumina MiSeq Amplicon sequencing SOP. Amplicons were purified using AMPure XP Beads (Beckman Coulter, Krefeld, Germany), and amplicon sequencing was then carried out in-house on a MiSeq Illumina platform (Illumina, Eindhoven, The Netherlands) with v3 chemistry.