Kit for BRAFV600E gene mutation and detection method
A kit and gene technology, applied in the fields of biotechnology and medicine, can solve the problem of high cost
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Embodiment 1
[0034] The method for detecting 7 kinds of gene mutations of BRAF V600E by real-time fluorescent PCR of the present invention comprises the following steps:
[0035] 1. Nucleic acid extraction
[0036] Taking paraffin-embedded tissue as an example, extract genomic DNA from paraffin-embedded tissue according to the recommended QIAamp DNA FFPE Tissue Kit instructions.
[0037] 2. Preparation of PCR reaction system (25ul)
[0038] Reagent name
Amount added
PCR reaction solution
16ul
Enzyme 1
2ul
Primer Probe Mix
2ul
template DNA
5ul
[0039] The template DNA includes sample DNA and negative and positive control DNA.
[0040] 3. PCR amplification
[0041] Real-time PCR amplification conditions are: 50°C for 2min, 1 cycle; 95°C for 5min; 95°C for 10s, 55°C for 15s, 72°C for 30s, 5 cycles; 95°C for 10s, 60°C for 45s (collect FAM and HEX fluorescence signals) , 40 cycles.
[0042] 4. Result judgment
[0043] 1) In additi...
Embodiment 2
[0051] The invention was used to detect 10 cases of tumor tissue samples from patients with non-small cell lung cancer, and compared with the sequencing results, the result: 2 cases were detected positive for V600E, and the coincidence rate with the sequencing method was 100%.
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