Combination of mutation sites for detection of type 2 diabetes susceptibility genes, detection primers and applications
A type 2 diabetes and mutation site technology, applied in the field of molecular biology, can solve problems such as high cost, long detection time, and low accuracy
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Embodiment 1
[0033] This example is used to illustrate the method of using the primer combination described in the present invention to detect the mutation site described in the present invention, and the specific steps are as follows:
[0034] 1. Peripheral blood extraction
[0035] Whole blood was frozen in EDTA anticoagulant tubes at -80°C, and DNA was extracted using the QIAAmpgDNABlood Minikit.
[0036] 1) Take 200 μL blood sample with added anticoagulant to a 1.5 mL centrifuge tube, then add 40 μL proteinase K solution,
[0037] Then add 200μL AL Buffer, and immediately vortex and oscillate thoroughly for 15 seconds;
[0038] 2) Incubate at 56°C for 30 minutes;
[0039] 3) Take out the cracked specimen from the oven and shake it shortly;
[0040] 4) Add 200 μL of absolute ethanol, immediately moisten and vortex to fully mix, and shake briefly;
[0041] 5) Add the mixture of the previous step (including possible precipitation) into an adsorption column (the adsorption column is pl...
Embodiment 2
[0094] In this example, using the method described in Example 1, 1054 diabetic patients were tested for genes, and 1028 were determined to be type 2 diabetic patients, with a detection accuracy rate of 97.5%. Further, for 512 Asian healthy persons data from the 1000G (Thousand Genomes Project) public database, the genotypes of the 21 mutation sites described in the present invention were analyzed, and the formula in Example 1 was used to calculate according to the analysis results, and judged 451 were non-type 2 diabetes patients, and the detection accuracy rate was 88.1%. The comprehensive detection accuracy rate is 92.8%,
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