Method for detecting hypertension susceptible gene and detection kit
A susceptibility gene and high blood pressure technology, applied in the fields of molecular biology and medicine, can solve the problems of unconfirmed correlation and few studies, and achieve the effects of simple diagnosis and treatment, low cost, and simple and easy methods
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Embodiment 1
[0048] Fluorescent PCR detection
[0049] 1. Experimental materials
[0050] The 7900HT fluorescent quantitative PCR instrument was purchased from ABI Company of the United States, and the polymerase chain reaction solution (TaqMan EXPressMaster Mix) was custom-synthesized by Applied Biosystems (ABI).
[0051] 2. Primer and probe design and synthesis
[0052] Using the partial sequence of intron 2 of the MFn2 gene as a template, the Primer ExpressTM2.0 software was used to analyze the TaqMan primers and probe sites, and custom-synthesized by Applied Biosystems (ABI).
[0053] Primers for detection:
[0054] MFn2 gene rs4240897 upstream primer sequence: 5′-GCTAGGAGTTGCTGTACATAAC-3′
[0055] MFn2 gene rs4240897 downstream primer sequence: 5′-AGGTGGTCAACAGCTTTGGCT-3′
[0056] Fluorescent probes:
[0057] MFn2 gene rs4240897 fluorescent probe 1: 5′-VIC-TACTCCTCAACAAATAG-TAMRA-3′
[0058] MFn2 gene rs4240897 fluorescent probe 2: 5′-FAM-TACTCCTCAACGAATAG-TAMRA-3′
[0059] 3. ...
Embodiment 2
[0073] The rs4240897 site of the essential hypertension susceptibility gene MFn2 gene was detected by sequencing. 10 cases of each of the above hypertension cases and control groups were selected for sequencing to determine the genotype of rs4240897.
[0074] 1. Experimental method
[0075] The above-mentioned fluorescent PCR primers were still used as primers for PCR sequencing, and the amplified products were directly sequenced after purification. The sequencing instrument is the 3130xl genetic analyzer of ABI Company, which is analyzed with sequence analysis 5.2 analysis software, and the results can also be viewed with chromas.
[0076] 2. Experimental results
[0077] The screenshot of the sequencing result is as follows: figure 1 shown. In the end, the sequencing results of 20 cases were completely consistent with the genotype analysis results of 7900 fluorescent PCR.
[0078] 3. Association analysis of MFn2 gene rs4240897 genotype and hypertension susceptibility
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