Kit and method for detecting polymorphism of CYP3A4 gene
A gene polymorphism and kit technology, applied in the direction of biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems of CYP3A4 gene polymorphism, such as long time, long time, cumbersome steps, etc.
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Embodiment 1
[0071] Embodiment 1. Preparation of kit
[0072] 1. Primer design for internal reference gene ACTB and CYP3A4 gene
[0073]According to the gene sequence, the ACTB gene sequence and the CYP3A4 gene sequence are derived from the National Center for Biotechnology Information Nucleic Acid Database (NCBI), wherein the ACTB gene ID is 60, and the reference sequence number is NG_007992.1, also refer to the SEQ ID NO in the sequence list of the present invention :35; the CYP3A4 gene ID is 1574, and the reference sequence number is NG_008421.1. Primer5.0 primer design software was used to design specific primers for the internal reference gene ACTB and the CYP3A4 gene, including: the long fragment of the 1st to 3rd exons of the CYP3A4 gene Amplification-specific upstream and downstream primers, long fragment amplification specific upstream and downstream primers of exons 4 to 7 of CYP3A4 gene, long fragment amplification specific upstream and downstream primers of exons 8 to 11 of CYP...
Embodiment 2
[0122] Example 2. Detection of CYP3A4 gene polymorphism with the kit prepared in Example 1
[0123] Take the results of random detection of peripheral blood samples from 30 subjects as an example.
[0124] The detection process of using the kit of the present invention to detect the CYP3A4 gene polymorphism in the population is as follows: first obtain the peripheral blood sample of the clinical subject, and quickly extract the genomic DNA; secondly, first prepare the PCR reaction solution of the internal reference gene ACTB, and then add the concentration of 2 ng / μl of internal reference gene ACTB sequence and 2 μl of internal positive control sequence were used for PCR amplification. After PCR, electrophoresis was performed in agarose DNA gel with a concentration of 2% to check the PCR amplification. If there are two bands of about 390bp and 800bp in the PCR amplification product of the internal reference gene ACTB, it indicates that the entire detection process is effective...
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