Kit for genetic detection of thyroid cancer
A thyroid cancer and kit technology, which is used in the determination/inspection of microorganisms, material excitation analysis, fluorescence/phosphorescence, etc., can solve problems such as protein function changes, increased risk of thyroid cancer, and mismatch repair pathways cannot be performed normally.
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Embodiment 1
[0029] Example 1. Use of detection kits
[0030] Step 1: Extraction of DNA template
[0031] Genomic DNA of oral epithelial cells was extracted by silica gel adsorption.
[0032] Step 2: Real-time quantitative PCR reaction
[0033] Use the fluorescent quantitative PCR kit in the detection kit to perform 4 independent fluorescent quantitative PCR reactions. The total volume of each reaction is 10 μl, including 2 μl of DNA template with a concentration of 20 ng / μl, 1 μl of 10 X fluorescent quantitative PCR reaction buffer, 0.1μl 25mM dNTP mix, 0.6μl 25mM MgCl 2 solution, 0.025 μl (5 units / μl) Taq DNA polymerase, 0.225 μl each of 20 μM sense primer and antisense primer, 0.25 μl each of 10 μM fluorescent probe with VIC and fluorescent probe with FAM, and 5.325 μl of deionized water.
[0034] The reaction was carried out on a PCR amplification instrument, and the reaction conditions were 50° C. for 2 minutes, 95° C. for 10 minutes, and 60 cycles of 95° C. for 30 seconds and 60° ...
Embodiment 2
[0038] Example 2. Application of detection kits for individual thyroid cancer genetic susceptibility testing services
[0039] Step 1: DNA Extraction
[0040] The physicians in the laboratory department of the hospital instructed the subjects to use oral swabs to sample oral epithelial cells, and the silica gel adsorption method was used to extract DNA from oral epithelial cells.
[0041] Step 2: Genotyping Assays
[0042] Using the kit provided by the invention, the Arg399Gln SNP site on the XRCC1 gene, the A4541G SNP site on the XRCC3 gene, the C-2578A SNP site on the VEGFA gene and the gIVS12-6T>C SNP on the hMSH2 gene of the genomic DNA of the subject The loci were detected by fluorescent quantitative PCR to determine the genotypes of these 4 SNP loci.
[0043] Step 3: Analysis of Individual Genetic Susceptibility to Thyroid Cancer
[0044] Through the analysis of the SNP genotype of the tested subjects, an analysis report sheet of the individual's genetic susceptibilit...
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