Kit and method for detecting gene polymorphism capable of influencing mercaptopurine personalized medications by means of pyro sequencing method
A pyrosequencing method and gene polymorphism technology, applied in the field of molecular biology, can solve problems such as lack of activity, and achieve the effect of simple operation and small sample size
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[0026] TPMT (rs1142345) upstream primer: 5′- TGG GGA ATT GAC TGT CTT TTT GA-3′ (SEQ ID NO.2);
[0027] TPMT (rs1142345) downstream primer: 5′- TCC ATT ACA TTT TCA GGC TTT AGC-3′ (SEQ ID NO.3);
[0028] TPMT (rs1142345) sequencing primer: 5′- GAC TGT CTT TTT GAA AAG TT -3′ (SEQ ID NO.4);
[0029] 1. DNA extraction
[0030] 1.1 The preparation and inspection of reagent materials before the experiment are as follows:
[0031] (1) Check the shelf life of the kit and ensure that ethanol has been added to Wash Buffer 1 and 2, and tick the corresponding marks on the bottle; (2) Isopropanol (if not available, absolute ethanol can be used instead) and 75% ethanol ; (3) 1.5mL Eppendorf tubes and various pipette tips within the validity period of autoclaving.
[0032] 1.2 Take out the EDTA anticoagulant tube containing whole blood from the refrigerator at 4°C, and mix it upside down several times;
[0033] 1.3 Make a mark on the 1.5mL Eppendorf tube corresponding to the unique identi...
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