Gt 96 96 dynamic array

DNA extracted from salivary samples of IBS patients and HCs was processed and assessed by the UCLA Biological Samples Processing Core using the Autopure LS Nucleic Acid Purification instrument (Gentra Systems, Inc., Minneapolis, MN.) The COMT polymorphism (rs174697) was genotyped using a 5′ nuclease assay to discriminate between the two alleles A/G (Taqman SNP Genotyping Assay C_2255328_10, Applied Biosystems Inc.). Polymerase chain reactions were performed using 5-μL reaction volumes in 384-well plates with 5 ng of DNA. The standard protocol provided with the kit was followed. End point reads of fluorescence levels were obtained with an ABI 7900HT Sequence Detection System. The SNPType genotyping was done on the other 10 SNPs using the Fluidigm Biomark system. First, 10–60 ng of DNA was pre amplified using Qiagen Multiple PCR master mix. This was then diluted and used for amplification as starting material. Next, the samples and assays were loaded onto GT 96*96 Dynamic array and processed per Fluidigm protocol. The assays used were designed using Fluidigm’s proprietary technology. The genotyping calls were made using Fluidigm SNP genotyping software. The SNPs rs4680 and rs6280 were genotyped with the KASP genotyping system (KBiosciences, Ltd, Hoddesdon, UK) as recommended by the manufacturer using customized assays as outlined previously.[34 (link)]