16S Metabarcoding of Airborne Microbes in a Medical Center
The 16S rRNA gene from airborne bacterial samples was amplified using a thermal cycler with specific primers and PCR conditions. A 25 L reaction mixture was prepared, including primers, Terra PCR Direct buffer, polymerase, water, and the DNA sample. After amplification, the PCR products were purified using a size-selective purification kit. Indexation was performed with Illumina Nextera XT primers, and the amplicon size (~630 bp) was verified using an Agilent TapeStation. Purified amplicons were quantified using a Qubit fluorometer, and the 16S rRNA libraries were diluted and sequenced on an Illumina MiSeq System at the Universidad de La Frontera.