title: DNA-seq of human skin microbiome: bacteria
design: Isolation: ZymoBIOMICS DNA Miniprep Kit (Zymo Research); PCR: ITS1-5.8Srv and ITS1-18Sfw for fungal ITS1 region, KAPA HIFI HotStart ReadyMix (Roche), 1.5 M primers, 5 l of template, denaturation 95C 3 min, 35 cycles (95C 30s, 55C 30s, 72C 30s), elongation step 72C 5min using 5 ng/L DNA, in triplicates; Normalization: SequalPrep Normalization Plate Kit (ThermoFisher Scientific); Concentration increase in vacuum concentrator; Purification: DNA Clean & Concentrator Kit (Zymo Research); Adapters: TruSeq DNA Single Indexes Set A (Illumina) compatible with Miseq platform, ligated with KAPA HyperPrep kit (Roche); Sequencing: Miseq platform (Illumina) at CEITEC Genomics Core Facility (Brno, Czech Republic)