16S rRNA amplicons from grassland soils affected by cattle manure with or without additional chlortetracycline
AMPLICON: Barcoded PCR primers (926f/630r) targeting the V6-9 regions of small ribosomal subunit rRNA genes were used to construct libraries. Fusion primers were designed according to the guidelines of Roche (www.my454.com). The PCR reaction set up and amplification was performed according to (Chroтбkovб et al., 2015). Quality of amplicons was checked by agarose (2%) gel electrophoresis and quantified using a Quant-iT dsDNA BR assay kit (Invitrogen, CA, USA). Amplicons were pooled equimolar to create five forward-oriented libraries.