METHOD FOR DETERMINING DNA METHYLATION LEVEL AT SPECIFIC SITE IN BIOLOGICAL SAMPLE AND APPLICATION THEREOF

摘要

A method for determining a DNA methylation level at a specific site in a biological sample and an application thereof. The method comprises steps of: acquiring a biological sample and extracting a DNA fragment to be tested from the biological sample; modifying the DNA fragment to be tested to deaminize unmethylated cytosine in the DNA fragment so as to convert the cytosine into uracil; using the modified DNA fragment as a template DNA fragment, and using a first primer and a second primer to perform PCR amplification on the template DNA fragment from both sides of the template DNA fragment, respectively; processing the PCR product amplified by PCR to remove excessive first primers, second primers, and dNTPs, and performing purification and recovery; detecting methylation and demethylation fluorescence polarization values of a CpG site of the DNA fragment; and calculating, according to the methylation and demethylation fluorescence polarization values of the CpG site of the DNA fragment to be tested, the methylation level of the DNA fragment.