Individual genetic test reagent kit of medicines for relieving fever and alleviating pain
A gene detection, antipyretic and analgesic technology, applied in the biological field, can solve the problems affecting the treatment effect, toxic and side effects, etc.
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Embodiment 1
[0029] Prepare a kit for detecting the metabolism of antipyretic and analgesic drugs for one person, including the following components:
[0030] (1) 4 pairs of CYP2D6*10, CYP2C9*3, CYP2C19*2 / *3 gene PCR amplification and sequencing primers, 1 OD for each primer, see Table 1 for primer sequences;
[0031] (2) PCR amplification reagent: 7.5 µL of Taq enzyme mixture (dNTP, l0× PCR reaction buffer, MgCl 2 );
[0032] (3) PCR product purification reagent: 1.6 µL SAP enzyme mixture (SAP enzyme, ExoI enzyme, deionized water);
[0033] (3) Sequencing reagents: 2.2 µL BigDye mix (Bigdye, 5×seq), 2.5 µL EDTA, 30 µL ethanol solution (100%), 100 µL ethanol solution (70%), 8 µL Hi-Di.
[0034] This kit is stored at -20°C, and repeated freezing and thawing should be avoided as much as possible.
Embodiment 2
[0036] The steps to use the kit for detecting the metabolic capacity of antipyretic and analgesic drugs for one person include:
[0037] (1) Extract the genomic DNA of the sample;
[0038] (2) PCR amplification of CYP2D6*10, CYP2C9*3, CYP2C19*2 / *3 genes
[0039] Design primers for CYP2D6*10, CYP2C9*3, and CYP2C19*2 / *3 genes according to Table 2 for PCR amplification. Each reaction system has a total volume of 50ul, including 25µL of 2*Mix mixture, 17µL of deionized water, upstream and downstream 3 µL of each primer, 2 ul of genomic DNA; the PCR reaction conditions were 95°C for 3 minutes for pre-denaturation, followed by 35 cycles of 95°C for 30 sec, 60°C for 20 sec, and 72°C for 30 sec; and finally extension at 72°C for 5 minutes;
[0040] (3) Purification of PCR products
[0041] Each reaction system has a total volume of 5.6 µL, 4 µL of PCR product, and 1.6 µL of SAP enzyme mixture (SAP enzyme, ExoI enzyme, deionized water). The reaction conditions are 37°C for 45 minute...
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