U-TOP technology is a highly sensitive genotyping technology that enables to detect the genetic mutation (SNP, SNV, In/del, point mutation), even a single nucleotide sequence variation, based on a PNA probe using a Real-time PCR.
U-TOP technology integrates Ct analysis, which is commonly used in Real-Time PCR Diagnosis, and Melting Array analysis, which determines specific temperature changes in base sequences, to identify up to 16 genetic mutations only by one reaction. For example, if the combined normal binding temperature in DNA is 68℃ when one of the base sequences is mutated, the combined binding temperature drops to 58℃. These temperature differences allow us to identify each gene mutation.
DNA methylation test is divided into two major steps. First, a chemical called Bisulfite is used to distinguish methylated and unmethylated cytosine. The unmethylated cytosine converted to Uracil by Bisulfite, and methylated cytosine (C-CH3) remains as cytosine. The methylation status is then identified by the methylation-specific PCR method or base sequencing analysis called Pyrosequencing. Unmethylated DNA is not amplified due to the genetic modification by Bisulfite, but DNA methylated genes are amplified.