title: Illumina of cow milk bacteria
design: A 464-nucleotide sequence of the V3-V4 region of the 16S rRNA gene (Escherichia coli positions 341 to 805) was amplified starting from DNA. Unique barcodes were attached before the forward primers. The amplicons were cleaned using the Agencourt AMPure kit (Beckman coulter). DNA concentrations of the amplicons were determined using the Quant-iT PicoGreen dsDNA kit (Invitrogen). The quality of the generated amplicon libraries was evaluated by a Bioanalyzer 2100 (Agilent, Palo Alto, CA, USA) using the High Sensitivity DNA Kit (Agilent).