Human CYP2C19 gene polymorphism detection specific primer and kit
A CYP2C19, gene polymorphism technology, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., can solve the problem of unsatisfactory gene polymorphism detection effect, etc., to achieve true and credible results , Reduce pollution and stabilize the effect of typing detection
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Embodiment 1
[0055] Preparation of human CYP2C19 gene polymorphism detection kit of the present invention comprises the following steps:
[0056] 1. Primer and probe synthesis:
[0057] Design and synthesize three sets of specific primers: SEQ ID NO:1, SEQ ID NO:2 and SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5 and SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8 and SEQ ID NO:9; 3 sets of specific probes SEQ ID NO:10 and SEQ ID NO:11, SEQ ID NO:12 and SEQ ID NO:13, SEQ ID NO:14 and SEQ ID NO:15, and label the FAM fluorescent group at the 5' end of SEQ ID NO:10, SEQ ID NO:12 and SEQ ID NO:14, and label the NFQ-MGB non-luminescent quenching group at the 3' end, in SEQ ID NO:14 The 5' end of NO:11, SEQ ID NO:13 and SEQ ID NO:15 is labeled with VIC fluorescent group, and the 3' end is labeled with NFQ-MGB non-emission quenching group. The primers and probes were prepared into 100 μM stock solutions for storage.
[0058] 2. Prepare the internal standard system: design and synthesize a pair of internal stand...
Embodiment 2
[0071] The human CYP2C19 gene polymorphism detection kit prepared in Example 1 was used to detect the samples to be tested.
[0072] In this embodiment, the anticoagulated whole blood samples of 40 cardiovascular patients were collected, and genomic DNA was extracted therefrom, and the CYP2C19 gene polymorphism of the sample to be tested was detected with the human CYP2C19 gene polymorphism detection kit obtained in Example 1 Condition.
[0073] 1. Genomic DNA extraction from blood samples
[0074]Take 300 μl of whole blood, add 900 μl of cell lysate CL, invert and mix well, let stand for 5 minutes, centrifuge at 10,000 rpm (11,500×g) for 1 minute, absorb the supernatant, leave the cell pellet, add to the cell pellet collected by centrifugation 200μl Buffer GS, shake until thoroughly mixed. Add 20 μl Proteinase K solution and mix well. Add 200 μl buffer GB, mix well by inversion, place at 56°C for 10 minutes, invert and mix several times during this time, the solution shoul...
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