title: NICU room sample
design: Genomic DNA from room samples were subjected to 16S rRNA V3-4 MiSeq library preparation which included dual-barcoded multiplexing with a heterogeneity spacer for higher sequence quality [Fadrosh et al, Microbiome 2014]. Two microliters of 5X concentrated gDNA template was used in the reaction and run at 35 cycles. Amplicons were purified using the Just-a-Plate PCR normalization and purification kit (Charm Biotech, San Diego, CA, USA). Equal amounts of each sample were sent to the University of California Davis DNA Technologies Core Facility (http://dnatech.genomecenter.ucdavis.edu) and run on a MiSeq with v3 300PE chemistry.