Reagent kit detecting pulmonary cancer susceptibility through CYP2E1 gene
A kit and susceptibility technology, which is applied in the field of kits for detecting susceptibility to lung cancer, and can solve problems such as the decrease of CYP2E1 transcription level
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Embodiment 1
[0023] Example 1. Use of detection kits
[0024] Step 1: Extraction of DNA template
[0025] Genomic DNA of oral epithelial cells was extracted by silica gel adsorption.
[0026] Step 2: Real-time quantitative PCR reaction
[0027] Use a fluorescent quantitative PCR detection kit that can detect susceptibility to lung cancer, which contains the following primer pairs and fluorescent probes:
[0028] Sense primer: 5'-ACAAGTGATTTGGCTGGATT-3' (SEQ ID NO: 2) with a Tm value of 56°C
[0029] Antisense primer: 5'-AATGAAGAGAATGTTTTTCATT-3' (SEQ ID NO: 3) with a Tm value of 54°C
[0030] Fluorescent probe: 5'-AGATTCATTGTTAATAATAAAAGTATAAA-3' (SEQ ID NO: 4) with a Tm value of 64°C
[0031] The fluorescent quantitative PCR reaction system has a total volume of 10 μl, including 2 μl of DNA template with a concentration of 20 ng / μl, 1 μl of 10X fluorescent quantitative PCR reaction buffer, 0.1 μl of 25mM dNTP mixture, and 0.6 μl of 25mM MgCl 2 solution, 0.025 μl (5 units / μl) Taq DNA ...
Embodiment 2
[0035] Example 2. Services that guide people to proactively prevent lung cancer
[0036] At present, Shanghai Zhujian Bioengineering Co., Ltd. has launched this service.
[0037] Step 1: DNA Extraction
[0038] The subjects were consulted before the service, and the subjects signed the informed consent and filled out the questionnaire on living and eating habits. According to the instructions in the sampling box, the oral epithelial cells were sampled with oral swabs, and the DNA of the oral epithelial cells was extracted by silica gel adsorption.
[0039] Step 2: Genotyping Assays
[0040] Using the kit provided by the present invention, the SNP site rs2031920 on the CYP2E1 gene of the tested subject's DNA is detected by fluorescent quantitative PCR, and the genotype of the SNP site is determined.
[0041] Step 3: Instruct people to be proactive in preventing lung cancer
[0042] Through the analysis of the SNP genotype of the tested person, the genotyping test report and...
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