16S amplicon (microbiome) of mouse spleen
DNA templates were processed for the 16S rRNA gene amplification according to Earth Microbiome Project standards (EMP; http://www.earthmicrobiome.org/protocols-and-standards/16s/). Samples were multiplexed based on a double barcoding strategy with 12-bp Golay barcodes (EMP protocol) combined with the forward primer 515F (Parada et al 2016) and 926R primers (Parada et al 2016, Walters et al 2016) yielding 450p long amplicons of V4 hypervariable region of the 16S rRNA gene. The library was sequenced on the single MiSeq run using v2 chemistry.