Isolation of microbial DNA from BALF, 16S and ITS2 metabarcoding, and sequencing of the amplicon pools
Microbial DNA was isolated from (1–2 mL) of BALF supernatant using our previously published protocol, which was specifically designed to improve the recovery of microbial DNA from low biomass BALF samples [To identify and characterize the fungal population associated with the BALF specimens, we carried out fungal DNA barcoding using the internal transcribed spacer (ITS) 2 region. Amplicon libraries were constructed at Genewiz (China) using forward and reverse primers containing the sequences 