For 16S rRNA sequencing, single colonies were suspended in 100 μl of sterile water and heated to 100°C for 15 min. The suspension was centrifuged for 10 s at 6,400 rpm and 5 μl of each suspension were used as the template for PCR amplification of the 16S rRNA gene. PCR was performed using a mix of 25 μl of GoTaq® Green Master Mix (Promega, Madison, WI, USA), 18 μl of PCR certified water (Promega), and 1 μl each of forward primer (8FPL, AGTTTGATCCTGGCTCAG) and reverse primer (806R, GGACTACCAGGGTATCTAAT) as previously described (Vornhagen et al. 2013 (link)). PCR products were cleaned using the QIAquick PCR Purification System (Qiagen, Valencia, CA, USA) according to the manufacturer's protocol. The purified DNA was checked for quantity and purity using a NanoQuant Tecan M200 (Tecan, Durham, NC, USA). Sequencing was performed by the DNA Sequencing Core at the University of Michigan (Ann Arbor, MI, USA) using Applied Biosystems 3730xl DNA Analyzers (Applied Biosystems, Carlsbad, CA, USA), BigDyev3.1 chemistry (MCLAB, San Francisco, CA, USA), and the protocols recommended by the manufacturer. Resulting partial 16s rRNA gene sequences were analyzed using CHROMAS (Brisbane, Australia) and compared to known sequences in the National Center for Biotechnology (NCBI) database using the Basic Local Alignment Search Tool (BLAST).