Kit for detecting TERT gene promoter mutation, and detection method and application of kit
A kit and promoter technology, applied in the field of clinical medical gene detection, can solve the problems of complex reaction system of the kit, many influencing factors, heavy workload, etc., and achieve the effects of low price, few influencing factors, and accurate detection.
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[0049] Embodiment 1: Detection of TERT gene promoter mutation
[0050] The kit for detecting TERT gene promoter mutations is a method for detecting TERT gene promoter mutations in non-disease diagnosis, comprising the following steps:
[0051] (1) Extract the urine of the sample to be tested, centrifuge to extract the urine cells, extract the whole DNA of the cells, and use the specific primers of SEQ ID NO.3-4 to carry out real-time PCR amplification of the template DNA;
[0052] Upstream primer: 5'-cccttcaccttccagctc-3' (SEQ ID NO.3);
[0053] Downstream primer: 5'-agcgctgcctgaaactgg-3' (SEQ ID NO.4);
[0054] (2) adding the fragment amplified in step (1) to the fluorescent probe of SEQ ID NO.1-2 for heating and melting;
[0055] The nucleotide sequence of the donor probe is as follows:
[0056] Donor probe: 5'-CGTCCCGACCCCTCCCGGGTC-Flc-3' (SEQ ID NO.1);
[0057] Receptor probe: 5'-Lc640-GCCCAGCCCCTTCCGGGCCCT-Phos-3' (SEQ ID NO.2);
[0058] (3) The mutation of the TERT ...
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