16S rRNA Seq of sputum microbiome : Control
Sample procaryotic DNA was extracted using FastDNA Spin Kit For Soil (MP Biomedicals) based on the manufacturers recommendation. From each sample, 500 l of sputum was used for DNA extraction. The DNA concentration were monitored using Qubit dsDNA Assay Kit in Qubit Flurometer (Life Technologies,CA, USA). Fifty ng of each of the extracted and purified sputum DNAs was used for the following amplification of 16S rRNA genes. Amplification of 16S rRNA was carried out according to the Illumina protocol Preparing 16S Ribosomal RNA Gene Amplicons for the Illumina MiSeq System.