基于分子信标方法对DNAzyme反应的优化与检测

Optimization and detection of DNAzyme reaction based on molecular beacon method

中文关键词:  DNAzyme  分子信标  条件优化  定量检测

英文关键词:DNAzyme  molecular beacon  optimization  quantitative detection

中文摘要:

      目的 建立DNAzyme与分子信标结合–切割–释放的级联放大方法, 优化反应的条件, 对DNAzyme进行定量检测。方法 选择4个优化条件, 分别是: 金属离子种类、金属离子浓度、pH值和反应时间。在最佳条件下建立标准曲线, 对DNAzyme进行检测。结果 本方法的最佳反应条件为: Mg2+金属离子催化反应、Mg2+浓度为10 mmol/L、pH为9.5、反应时间为4 h。标准曲线为Y=17.1628X+420.7767(r2=0.9402), 检测范围为0.5~100 nmol/L。结论 此方法理论正确, 实验的准确度高, 为DNAzyme的应用提供了一个成熟的方法和新的思路。

英文摘要:

      Objective To establish a cascade amplification method combining DNAzyme with molecular beacon-cleavage-release, optimize the reaction conditions, and quantitatively detect DNAzyme. Methods Four optimization conditions were selected, including metal ion species, metal ion concentration, pH and reaction time. A standard curve was established under optimal conditions to detect DNAzymes. Results The optimal reaction conditions for this method were: Mg2+ metal ion catalytic reaction, Mg2+ concentration of 10 mmol/L, pH of 9.5, and reaction time of 4 h. The standard curve was Y=17.1628X+420.7767 (r2=0.9402) and the detection range was 0.5?100 nmol/L. Conclusion The method is correct and the accuracy of the experiment is high, , which provides a mature method and new ideas for the application of DNAzyme.