Gastrointestinal colonization of Candida albicans increased serum (1-->3)-beta-D-glucan, without candidemia, and worsened cecal ligation and puncture sepsis in murine model
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.