Detection method of electrochemical acute leukemia gene Pax-5a based on enzyme assisted cyclic signal amplification
A technology of acute leukemia and pax-5a, which is applied in the direction of biochemical equipment and methods, microbial measurement/inspection, etc., can solve problems such as limiting detection sensitivity, and achieve the effects of avoiding detection interference, improving reaction speed, and reducing complexity
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[0035] (1) Treat the thiol-modified probe (TP) with 2mM TCEP at 37°C for 2 hours to reduce the presence of disulfide bonds; then, add 10 μL of 0.3 μM treated thiol-modified probe (TP) solution dropwise To the surface of the gold electrode, incubate at 4°C for 12 hours, then rinse the electrode with Tris-HCl buffer (10mM, 1mM EDTA) to remove unbound TP probes, and block the electrode with 2mM MCH to obtain the treated modified Gold electrode MCH / TP / AuE;
[0036](2) Put 2 μL of hairpin probe (HP) with a concentration of 10 μM and 1 μL of Pax-5a standard aqueous solution with a concentration of 500pM, 100pM, 50pM, 10pM, 5pM, 1pM, 500fM, 100fM, 10fM, and 0fM in the reaction tube Mix in medium, add CutSmart buffer (20mM Tris-HAc, 50mM KAc, 10mM MgAc2, 0.1g / mL BSA, pH 7.9), incubate at 37°C for 1 hour; Add 5U / μL of Klenow Fragment, 1μL of 10mM dNTPs and 1μL of 10U / μL Nt.BbvCI, incubate at 37°C for 3 hours, then heat at 80°C for 20 minutes to inactivate the enzyme;
[0037] (3) Tra...
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