title: Gastrointestinal colonization of Candida albicans increased serum (1-->3)-beta-D-glucan, without candidemia, and worsened cecal ligation and puncture sepsis in murine model
design: The amplification of the 16S rRNA genes are performed according to Caparaso et al. (2012), using universal prokaryotic primers 515F (5'-GTGCCAGCMGCCGCGGTAA-3') and 806R (5'-GGACTACHVGGGTWTCTAAT-3'). The primers are appended 5' Illumina adapter and 3' Golay barcode sequences. Each 25-μl PCR reaction comprises 1x EmeraldAmp® GT PCR Master Mix (TaKaRa, Shiga, Japan), 0.2 μM of each primer, and 75 ng of the metagenomic DNA. The PCR conditions are 94°C for 3 minute, and 35 cycles of 94°C for 45 second, 50°C for 60 second and 72°C for 90 second, followed by 72°C for 10 min. Independently trireplicate PCRs are performed per samples, and pooled.