B-raf gene mutation detection kit
A kit and gene technology, applied in the field of gene mutation detection products, can solve the problems of high detection cost, unsuitable for widespread promotion, and high cost of kit detection, and achieve the effect of avoiding repeated experiments
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Embodiment 1
[0038] 1. The composition of the kit.
[0039] The kit for detecting B-raf gene mutation in this embodiment includes: PCR reaction solution, quality control primer probe internal standard mixed solution, detection primer probe internal standard mixed solution, weak positive control solution and blank control solution, As shown in Table 1. The PCR reaction solution was prepared from 10×PCR buffer, 2mM dNTPs and 5U / μl hot-start enzyme. 10×PCR buffer includes 100mM Tris-HCl, 500mM KCl and 15mM MgCl 2 , the pH value of the Tris-HCl buffer used to configure the PCR buffer is 8.3. 2mM dNTPs include 2mM dATP, 2mM dGTP, 2mM dCTP and 2mM dTTP, respectively. The hot start enzyme is Taq DNA polymerase. Ten times the concentration of PCR buffer and 2mM dNTPs are packed in tube A, quality control primer probe internal standard mixture is packed in tube B, detection primer probe internal standard mixture is packed in tube C, hot start enzyme, weak positive control Solution and blank con...
Embodiment 2
[0085] The remainder of the kit for detecting B-raf gene mutations in this embodiment is the same as in Example 1, except that the 5' end of the nucleotide sequence of the B-raf gene-specific probe is provided with a HEX fluorescent label , the 5' end of the nucleotide sequence of the internal standard-specific probe is provided with a FAM fluorescent label. During the PCR reaction, the HEX channel collects the fluorescent signal of the B-raf gene, and the FAM channel collects the fluorescent signal of the internal standard gene.
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