Label-free fluorescence aptamer sensor detection adenosine based on double-amplification strategy construction
An aptamer sensor, label-free technology, which can be used in the determination/inspection of microorganisms, biochemical equipment and methods, etc., and can solve the problems of affecting the binding of aptamers to the target and reducing the binding constant of the aptamer itself.
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[0069] Example 1 A label-free fluorescent aptasensor constructed based on a dual amplification strategy was used to detect adenosine.
[0070] 1. Materials and Reagents
[0071] All nucleic acid probes used in this example were synthesized and purified by Sangon Bioengineering (Shanghai) Co., Ltd., and the base sequences are shown in Table 1. The DNA stock solution was TE Buffer (10 mM Tris-HCl, pH 8.0, 1 mM EDTA-2Na), and it was stored at -20°C. Adenosine was purchased from Sigma-Aldrich (Shanghai, China). Cytidine, uridine, guanosine, Exo III and DMSO were all purchased from Shanghai Sangong. Streptavidinized magnetic spheres (350 nm in diameter, 0.05% Tween-20, and 10 μM EDTA at a concentration of 3.324×10 11 beads mL -1 ,1.343g mL -1), Tris (>99.8%) was purchased from Amresco Inc. NMM (N-methylmesoporphyrinIX) was purchased from J&K Scientific Ltd. (Beijing, China). The NMM stock solution was dissolved in DMSO and stored at -20°C in the dark. All reagents (analytic...
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