High-throughput metagenomic sequencing to analyze the V4 variable region of 16S rRNA within the environmental samples collected from a cardiac hospital setting.
The barcoded V4 amplicons from each sample were then pooled in equal amounts to construct the DNA library. The size and concentration of the DNA library fragment were determined using tape station and Kapa quantitative PCR (qPCR) (Kapa Biosystems, USA). The final DNA library (600 L of 6 pmol/L library) was loaded onto the MiSeq v2, 2  250 cycle cartridge (Illumina, USA) and sequenced using the Illumina MiSeq platform at the Interdisciplinary Center for Biotechnology Research (ICBR) at the University of Florida