Human CYP2C9 and VKORC1 genetic polymorphism detection kit
A gene polymorphism and detection kit technology, which can be used in the determination/inspection of microorganisms, biochemical equipment and methods, etc. The effect of stable typing detection
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Embodiment 1
[0071] Preparation of CYP2C9 and VKORC1 gene polymorphism detection kit of the present invention comprises the following steps:
[0072] 1. Primer and probe synthesis:
[0073] Design and synthesize 2 groups of specific primers SEQIDNO:1, SEQIDNO:2, SEQIDNO:3 and SEQIDNO:4; SEQIDNO:5 and SEQIDNO:6; 2 groups of specific probes SEQIDNO:7 and SEQIDNO:8, SEQIDNO:9 and SEQIDNO:10, and label FAM fluorescent group at the 5' end of SEQIDNO:7 and SEQIDNO:9, label NFQ-MGB non-luminescence quenching group at the 3' end, label at the 5' end of SEQIDNO:8 and SEQIDNO:10 VIC fluorescent group, 3' end labeled NFQ-MGB non-emission quenching group. The primers and probes were prepared into 100 μM stock solutions for storage.
[0074] 2. Prepare the internal standard system: design and synthesize a pair of internal standard primers for the human-derived gene GADPH, the primer pair sequences are SEQ ID NO: 11 and SEQ ID NO: 12; design and synthesize the internal standard probe, the probe is SE...
Embodiment 2
[0081] The CYP2C9 and VKORC1 gene polymorphism detection kit prepared in Example 1 was used to detect the samples to be tested.
[0082] In this embodiment, 40 cases of anticoagulated whole blood samples were collected, and genomic DNA was extracted from them, and the CYP2C9 and VKORC1 gene polymorphism detection kits obtained in Example 1 were used to detect the gene polymorphisms of CYP2C9 and VKORC1 in the samples to be tested .
[0083] 1. Genomic DNA extraction from blood samples
[0084] 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 ProteinaseK solution and mix well. Add 200 μl buffer GB, mix thoroughly by inversion, place at 56°C for 10 minutes, invert and mix several times until the solution becomes clear (i...
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