Taqman hydrolysis probe and method for quantitatively detecting methylation level of MGMT (O6-methylguanine-DNA-methyltransferase)
A methylation and non-methylation technology, used in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc. achieve a specific effect
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[0042] Aiming at the 154-172 DNA sequence of the human MGMT gene promoter region, the present invention designs a Taqman fluorescent hydrolysis probe, which is complementary to the DNA sequence of this segment. Its base composition is: GATTTGGTGAGTGTUTGGG, the probe can extend 5 bases upstream (that is, complementary to the DNA sequence of segment 149-172) or extend 5 bases downstream (that is, complementary to the DNA sequence of segment 154-177) . In combination with the primers for detecting MGMT methylation by the MSP method, fluorescent quantitative detection of MGMT gene methylation was performed using a fluorescent quantitative technique.
[0043] The principle of the detection process of the present invention is as follows: the DNA fragment to be detected is first treated with bisulfite. Design a probe that can be complementary to the site to be tested. The 5' end of the probe is connected to the reporter fluorescence, and the 3' end is connected to the quencher fluor...
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