Cfx96 touch real time polymerase chain reaction detection system

We analyzed the ITGAM rs1143679G/A (R77H), rs1143683C/T (A858V), rs34572943C/T, and rs9888739C/T SNPs, which were determined by an allelic discrimination assay and TaqMan probes in a CFX96 Touch^™ Real-Time Polymerase Chain Reaction (PCR) Detection System (Bio‐Rad, Hercules, California, USA), according to the manufacturer’s instructions. The PCR reaction for each individual contained 10 ng of DNA, 2.5 μl of TaqMan Master Mix (Applied Biosystems, Foster City, California, USA), 0.065 μl of 40× assay mixture (Applied Biosystems, Foster City, California, USA), and 2.435 μl of DNase free water in a final volume of 5 μl. The PCR conditions used for amplification were as follows: Pre-PCR (1 cycle) 50°C for 2 minutes and 95°C for 8 minutes, followed by 45 cycles of denaturing at 95°C for 15 seconds, and annealing and extension at 60°C for 1 minute. We genotyped 35% of the samples twice and the reproducibility was 100%.

A commercial fungal DNA isolation kit, YeaStar Genomic DNA Kit^TM (Cat. No.D2002), was purchased from ZYMO RESEARCH (Irvine, USA. A C-Chip used as disposable hemocytometer was purchased from INCYTO (Cheonan, Republic of Korea). Field-emission scanning electron microscopy (FE-SEM) on a JSM-7500F instrument (JEOL, Seoul, Republic of Korea) was used to confirm the reaction and decoration materials. Zeta potentials of materials were measured using dynamic light scattering (DLS) on a DynaPro NanoStar instrument (Wyatt, Malvern, United Kingdom). Vortex mixer (T5AL) and a mini manufacturer (LABOGENE 1730R) MSH-30d stirring heater produced by Daihan Scientific Co., Ltd (Wonju-Si, South Korea) were obtained. Magic mixer (TMM-5) used for rotating the mixture tube during incubation was purchased from TOPSCIEN Instrument Co., Ltd (Ningbo, China). A CFX96 touch real-time polymerase chain reaction (PCR) detection system was purchased from BIO-RAD (Hercules, USA).