Reagent kit for detecting heart brain disease genetic susceptibility
A kit, susceptibility technology, applied in the fields of molecular biology and medicine, can solve problems such as discomfort, decreased ability to metabolize acetaldehyde, alcoholism, etc.
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Embodiment 1
[0014] Example 1. Use of detection kits
[0015] Step 1: Extraction of DNA template
[0016] Genomic DNA of oral epithelial cells was extracted by silica gel adsorption.
[0017] Step 2: Real-time quantitative PCR reaction
[0018] Using the detection kit, an independent fluorescent quantitative PCR reaction was carried out for each SNP site. The total volume of each reaction was 10 μl, including 2 μl of DNA template with a concentration of 20 ng / μl, 1 μl of 10X fluorescent quantitative PCR reaction buffer, 0.1μl 25mM dNTP mixture, 0.6μl 25mM MgCl 2 solution, 0.025 μl (5 units / μl) Taq DNA polymerase, 0.225 μl each of 20 μM primers, 0.25 μl each of 10 μM fluorescent probes, and 5.325 μl of deionized water.
[0019] The reaction was carried out on an ABI9700 type PCR amplification instrument, and the reaction conditions were 50° C. for 2 minutes, 95° C. for 10 minutes, and 60 cycles of 95° C. for 30 seconds and 60° C. for 1 minute. After the reaction, the fluorescence of the...
Embodiment 2
[0022] Example 2. Services for detection of genetic susceptibility to heart and brain diseases
[0023] Step 1: DNA Extraction
[0024] The physicians in the laboratory department of the hospital instructed the subjects to use oral swabs to sample oral epithelial cells, and the silica gel adsorption method was used to extract DNA from oral epithelial cells.
[0025] Step 2: Genotyping Assays
[0026] Use the kit provided by the present invention to detect ABCA1, ACE, ADIPOQ, AGT, APOB, APOE, AT1R, CYP11B2, ENOS, LEP, LPL, MTHFR, PON1, PPARG, SOD3, UCP2 and VDR genes on the genomic DNA of the subject The 18 SNP sites were detected by fluorescent quantitative PCR to determine the genotypes of these SNP sites.
[0027] Step 3: Analysis of genetic susceptibility to heart and brain diseases
[0028] Through the analysis of the SNPs genotypes of the subjects, an analysis report of genetic susceptibility to heart and brain diseases is issued, and the doctor explains and interprets...
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