Method for detecting rs4846085 locus of hypertension susceptibility gene and detection kit thereof
A kit and technology for hypertension, applied in the fields of molecular biology and medicine, can solve the problems of unproven correlation and little research, and achieve the effects of simple diagnosis and treatment, low cost, and simple and easy method.
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Embodiment 1
[0033] Example 1 Fluorescent PCR detection
[0034] 1. Experimental materials
[0035] The 7900HT fluorescent quantitative PCR instrument was purchased from ABI Company of the United States, and the polymerase chain reaction solution (TaqMan EXPress MasterMix) was custom-synthesized by Applied Biosystems (ABI).
[0036] 2. Primer and probe design and synthesis:
[0037] Using the partial sequence of intron 3 of the MFn2 gene as a template, the Primer ExpressTM 2.0 software was used to analyze the TaqMan primers and probe sites, and custom-synthesized by Applied Biosystems (ABI).
[0038] Primers for detection:
[0039] MFn2 gene rs4846085 upstream primer sequence: 5'-TGCGTAATGCAGGTGTTTATG-3' (SEQ ID NO 2)
[0040] MFn2 gene rs4846085 downstream primer sequence: 5'-CTCTTGTTACCTGCCCGTTAG-3' (SEQ ID NO 3)
[0041] Fluorescent probes:
[0042] MFn2 gene rs4846085 fluorescent probe 1: 5'-VIC-TGTGATGCATGCTTTGA-TAMRA-3' (SEQ ID NO 4)
[0043] MFn2 gene rs4846085 fluorescent pro...
Embodiment 2
[0054] The rs4846085 locus 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 rs4846085.
[0055] 1. Experimental method
[0056] 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.
[0057] 2. Experimental results
[0058] In the end, the sequencing results of 20 cases were completely consistent with the genotype analysis results of 7900 fluorescent PCR.
[0059] 3. Association analysis of MFn2 gene rs4846085 genotype and susceptibility to hypertension
[0060] RxC χ2 test was used to compare the distribution of MFn2 gene rs4846085 in...
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