Kit for detecting polymorphism of human CYP2C9 and VKORC1 genes
A gene polymorphism and kit technology, applied in the determination/testing of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., can solve problems such as inability to type multiple tubes, complicated amplification procedures, and complicated operations
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Embodiment 1
[0192] Example 1: Composition of the kit
[0193] The composition of the kit of the present invention is as follows:
[0194] Table 1 Composition of the kit
[0195]
Embodiment 2
[0196] Example 2: Extraction of genomic DNA from human whole blood / peripheral blood samples
[0197] In this embodiment, human whole blood / peripheral blood samples are collected, and genomic DNA is extracted therefrom.
[0198] Genomic DNA was extracted from EDTA anticoagulated plasma and used as a template for PCR detection. Use Zhengzhou Antu Bioengineering Co., Ltd.'s pretreatment reagents and nucleic acid extraction and purification reagents (magnetic bead method), and proceed according to the instructions. The details are as follows:
[0199] 1. Sample pretreatment:
[0200] Frozen whole blood is pre-treated with red blood cell lysate:
[0201] ① Take 200μL of clinical freeze-thaw or hemolyzed whole blood sample, add 600μL of red blood cell lysate, gently invert and mix 10
[0202] Second, after the red blood cells are lysed, the solution should be clear and transparent. Centrifuge at 12000rpm for 3min, discard the supernatant;
[0203] ②Add 600μL 0.2mg / ml proteinase K, incubate at...
Embodiment 3
[0215] Example 3: Crude extraction of genomic DNA from human oral swab samples
[0216] In this embodiment, the human oral swab sample is simply processed, and the genomic DNA of the sample is roughly extracted.
[0217] 1. Take a clean oral swab, extend the sampling site of the swab head into the left and right sides of the sampled person’s mouth, scrape up and down 5 times, the oral epithelial exfoliated cells will adhere to the swab head, and swab it quickly Soak the seeds in 1ml of rinse solution (purified water or normal saline can also be substituted), rinse them up and down 10 times, discard the swab, and save the swab rinse solution for later use.
[0218] 2. Place the swab rinse solution on a metal bath at 90°C and heat for 5 minutes. After cooling to room temperature, centrifuge at 12000 rpm for 3 minutes. Take the supernatant to obtain the crudely extracted genomic DNA.
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