Detection kit for triple negative breast cancer metastasis susceptibility gene TNF-alpha mutation and detection method
A technology of triple-negative breast cancer and a detection kit, which is applied in the fields of molecular biology and genetic diagnosis, and can solve the problems of poor prognosis, high risk of triple-negative breast cancer metastasis, and no molecular biology and genetic diagnosis methods for TNBC patients , to achieve the effect of ingenious design, accurate and reliable results
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[0033] A triple-negative breast cancer metastasis-related susceptibility gene TNF-α mutation detection kit, its components and contents are as follows: including 1.0ul of 10×PCR reaction buffer, 25mMMgCl 2 Solution 0.5ul, dNTPMixture (2.5mM) 1.0ul, TaqDNA polymerase 0.25ul (5U / ul), specific primers F and R (10uM) each 0.25ul, sterilized distilled water 6.0ul. This kit is used for one person-time detection, and the storage temperature is -20°C.
[0034] Step 1: DNA extraction from peripheral blood of patients with triple-negative breast cancer:
[0035] Genomic DNA from peripheral blood of patients was extracted using a blood genomic DNA extraction system (spin column method).
[0036] Step 2: PCR reaction, copy the target fragment:
[0037] (1) Design specific primers according to the gene sequence of the TNF-α promoter region to amplify the DNA fragment containing the rs1800629 site. The designed primer sequences are as follows:
[0038] F:5'-CCCCAAAAGAAATGGAGGCAATAGGTTTTG...
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