Reagent case for lung carcinoma susceptibility detection through PARP1 gene
A kit and susceptibility technology, applied in the field of kits for detecting lung cancer susceptibility through PARP1 gene, can solve the problems of reducing enzyme activity and increasing the risk of occurrence
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Embodiment 1
[0023] Example 1. Use of detection kit
[0024] Step 1: Extraction of DNA template
[0025] The genomic DNA of oral epithelial cells was extracted by silica gel adsorption method.
[0026] Step 2: Fluorescence quantitative PCR reaction
[0027] Use a fluorescent quantitative PCR detection kit that can detect lung cancer susceptibility, which contains the following primer pairs and fluorescent probes:
[0028] Sense primer: 5'-TTTGCCATTCACTGTGTTGG-3' (SEQ ID NO: 2) Tm value is 58°C
[0029] Antisense primer: 5'-ATCCTTGCTGCTATCATCA-3' (SEQ ID NO: 3) Tm value is 54°C
[0030] Fluorescent probe: 5'-CAGGCCAAGGCGGAAATGCT-3' (SEQ ID NO: 4) Tm value is 64°C
[0031] The fluorescence quantitative PCR reaction system is a total volume of 10μl, containing 2μl of DNA template at a concentration of 20ng / μl, 1μl of 10X fluorescence quantitative PCR reaction buffer, 0.1μl of 25mM dNTP mixture, 0.6μl of 25mM MgCl 2 Solution, 0.025μl (5units / μl) Taq DNA polymerase, 20μM sense primer and antisense ...
Embodiment 2
[0035] Example 2. Services to guide people to take the initiative to prevent lung cancer
[0036] At present, this service has been carried out in Shanghai Zhujian Biological Engineering Co., Ltd.
[0037] Step 1: DNA extraction
[0038] Conduct pre-service consultations for the tested persons, who will sign an informed consent form and fill in a questionnaire about eating habits. Use the oral sampling swab to sample oral epithelial cells according to the instructions in the sampling box, and use the silica gel adsorption method for DNA extraction of oral epithelial cells.
[0039] Step 2: Genotyping testing
[0040] Using the kit provided by the present invention, the rs1136410 SNP site on the PARP1 gene of the DNA of the testee is detected by fluorescent quantitative PCR to determine the genotype of the SNP site.
[0041] Step 3: Instruct people to take the initiative to prevent lung cancer
[0042] Through the analysis of the SNP genotype of the tested person, a genotyping test r...
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