Use and detection kit of locus rs2336384 of susceptibility gene of hypertension
A technology of susceptibility genes and kits, 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
[0034] Fluorescent PCR detection
[0035] 1. Experimental materials
[0036] The 7900HT fluorescent quantitative PCR instrument was purchased from ABI Company of the United States, and the polymerase chain reaction liquid (TaqMan EXPress Master Mix) was custom-synthesized by Applied Biosystems (ABI).
[0037] 2. Primer and probe design and synthesis:
[0038] Using the partial sequence of intron 1 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).
[0039] Primers for detection:
[0040] MFn2 gene rs2336384 upstream primer sequence: 5'-CTCAGAGCTGTACTGTCTTTAAG-3' (SEQ ID NO 2)
[0041] MFn2 gene rs2336384 downstream primer sequence: 5'-AGACAAGGGAAGGAAGTGAC-3' (SEQ ID NO 3)
[0042] Fluorescent probes:
[0043] MFn2 gene rs2336384 fluorescent probe 1: 5'-VIC-GCTGTGGCGTCATGCT-TAMRA-3' (SEQ ID NO 4)
[0044] MFn2 gene rs2336384 fluorescent probe 2: 5'-FA...
Embodiment 2
[0057] The rs2336384 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 rs2336384.
[0058] 1. Experimental method
[0059] 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.
[0060] 2. Experimental results
[0061] The screenshot of the sequencing result is as follows: figure 2 shown.
[0062] It can be seen that the sequencing results of 20 cases are completely consistent with the genotype analysis results of 7900 fluorescent PCR.
[0063] 3. Association analysis of MFn2 gene rs2336384 genotype and hypertension susceptibilit...
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