MTHFR, MTRR and RFC1 gene polymorphism detection primer combination and kit and application of MTHFR, MTRR and RFC1 gene polymorphism detection kit
A gene polymorphism and detection kit technology, which is applied in the direction of recombinant DNA technology, microbial measurement/inspection, biochemical equipment and methods, etc., can solve the problems of high cost of superimposition, high cost of technology, difficult conditions to control, etc., to achieve The effect of solving the problem of easy pollution, improving the detection efficiency and saving the detection cost
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specific Embodiment 2
[0056] A kind of detection method utilizing MTHFR, MTRR and RFC1 gene polymorphism detection kit of the present invention, concrete steps are as follows:
[0057] 1. Production of MTHFR, MTRR and RFC1 gene polymorphism detection kits based on the GeXP multiple gene expression genetic analysis system. The components included in the kit are the same as those in Example 1 above;
[0058] 2. Collection and extraction of DNA samples
[0059] Put the buccal swab scraped from oral epithelial cells into 300 μL DNA lysis buffer, treat it in a constant temperature mixer at 95°C and 1000 rpm for 5 minutes, take it out, leave it at room temperature until it cools down, and then add it to the sample 30 μL extraction buffer, mix well, centrifuge at 12000g for 5 minutes, the supernatant obtained is the DNA sample template for PCR;
[0060] 3. Use the extracted nucleic acid as a template for PCR reaction
[0061] 1) Add reagents and samples (PCR plate, see Table 4) to the 96-well sample pla...
specific Embodiment 3
[0083] Detection Kit Sensitivity and Specificity Analysis
[0084] Sensitivity analysis: After diluting the positive control substance according to a certain copy number ratio, it is detected by PCR amplification and capillary electrophoresis until no signal is detected. The copy number is the lowest detection line, which is the sensitivity of the kit. The kit has a sensitivity of 50 copies.
[0085] Specificity analysis: Single-plex PCR amplification is detected as a single peak of the target fragment size by capillary electrophoresis.
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