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Detection method for CYP4F2 gene polymorphism, as well as nucleic acid probe and kit for method
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A CYP4F2, gene polymorphism technology, used in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc.
Inactive Publication Date: 2015-04-29
FUWAI HOSPITAL OF CARDIOVASCULAR DESEASE CHINESE ACAD OF MEDICAL SCI
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Problems solved by technology
At present, CYP4F2 gene detection has been commercialized, but domestic medical institutions can only use western commercial products for warfarin sensitive gene detection
Method used
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Embodiment 1
[0108] 1. Preparation of probes and primer pairs for CYP4F2 gene
[0109] A probe for the CYP4F2 gene and a primer pair for the CYP4F2 gene were prepared using conventional techniques.
[0124] Aliquot, 19 μl per tube into 0.1ml PCR reaction tubes / plates, and transfer to the sample p...
Embodiment 2
[0138] Steps one to four:
[0139] In the same method and steps as in Example 1, the DNA sample was replaced by a CYP4F2 mutantDNA sample, and 1 μl of the mutantDNA sample was added to 19 μl of the PCR reaction solution.
[0140] 5. Results analysis
[0141] CYP4F2 appears to be mutant as a result of Figure 2 show. These graphs are analysis graphs showing changes in fluorescence intensity over time, that is, changes in fluorescence intensity as the number of amplification cycles increases. Such as Figure 2 As shown, the fluorescence curve indicates that the CYP4F2 gene in the sample is a mutant type.
Embodiment 3
[0143] Steps one to four:
[0144] In the same method and steps as in Example 1, the DNA sample was replaced with a CYP4F2 wild-type DNA sample, and 1 μl of the wild-type DNA sample was added to 19 μl of the PCR reaction solution.
[0145] 5. Results analysis
[0146] CYP4F2 expresses wild-type results in Figure 3 show. These graphs are analysis graphs showing changes in fluorescence intensity over time, that is, changes in fluorescence intensity as the number of amplification cycles increases. Such as Figure 3 As shown, the fluorescence curve indicates that the CYP4F2 gene in the sample is wild type.
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Abstract
The invention discloses a detecting probe for CYP4F2 gene polymorphism. The nucleotide sequences of the detecting probe are selected from a sequence 1 and a sequence 2; a fluorescent group and a quenching group are respectively arranged at the ends (5' and 3'); the peak values of the wavelengths of the florescent light emitted from the fluorescent group are at different positions. Preferably, the quenching group is selected from MGB and BHQ2; preferably, the fluorescent group is selected from FAM, VIC, JOE, HEX, CY3, NED, TAMRA, ROX, TEXAS RED, CY5 and the like. The invention further discloses a primer pair used for gene amplification; the primer pair comprises a forward primer and a reverse primer; the forward primer is an oligonucleotide which consists of base sequences of a sequence 3; the reverse primer is an oligonucleotide which consists of base sequences of a sequence 4. The invention further discloses a kit which contains the primers and the probe, and a detecting method for the CYP4F2 gene polymorphism.
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Application Information
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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6888C12Q2600/106C12Q2600/156
Inventor 李一石刘红韩璐璐田蕾娄莹王巍许建屏
Owner FUWAI HOSPITAL OF CARDIOVASCULAR DESEASE CHINESE ACAD OF MEDICAL SCI
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