title: ITS1,2 amplicon sequencing of lung sputum bacteria in humans: 17
design: DNA was extracted from samples using the DNEasy PowerSoil Pro Kit (Qiagen) according to the manufacturer's protocol. Barcoded PCR primers annealing to the 1 and 2 region of fungal internal transcribed spacer (Fwd, 5'-CTTGGTCATTTAGAGGAAGTAA-3'; Rev, 5'-GCTGCGTTCTTCATCGATGC-3') were used for library generation. The resulting library was sequenced on the Illumina MiSeq using 2x300 bp chemistry.