Method for detecting hypertension susceptibility 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
[0028] Embodiment 1 Fluorescence PCR detects
[0029] 1. Experimental materials
[0030] The 7900HT fluorescence 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).
[0031] 2. Primer and probe design and synthesis:
[0032] Using the partial sequence of intron 2 of the MFn2 gene as a template, TaqMan primers and probe sites were analyzed using Primer ExpressTM 2.0 software, and custom-synthesized by Applied Biosystems (ABI).
[0033] Primers for detection:
[0034] MFn2 gene rs 1474868 upstream primer sequence: 5'-TGAACTCCCAGCAGACCTTGTA-3' (SEQ ID NO 2)
[0035] MFn2 gene rs1474868 downstream primer sequence: 5'-GGTGCTTTTAAGTTCTGGTCAATGT-3' (SEQ ID NO 3)
[0036] Fluorescent probes:
[0037] MFn2 gene rs1474868 fluorescent probe 1: 5'-VIC-TGCAAGACTCGACAGC-TAMRA-3' (SEQ ID NO 4)
[0038] MFn2 gene rs1474868 fluorescent pro...
Embodiment 2
[0049] The rs1474868 site of the essential hypertension susceptibility gene MFn2 gene was detected by sequencing method. 10 cases of each of the above hypertension cases and control groups were selected for sequencing to determine the genotype of rs1474868.
[0050] 1. Experimental method
[0051] 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 ABI's 3130xl genetic analyzer, which is analyzed with sequence analysis 5.2 analysis software, and the results can also be viewed with chromas.
[0052] 2. Experimental results
[0053] In the end, the sequencing results of 20 cases were completely consistent with the genotype analysis results of 7900 fluorescent PCR.
[0054] 3. Association analysis of MFn2 gene rs1474868 genotype and hypertension susceptibility
[0055] Comparison of the distribution of MFn2 gene rs1474868 in hypertensive p...
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