马粪海胆RAPD反应体系的优化

Optimization of RAPD reaction system of Hemicentrotus pulcherrimus

以马粪海胆为研究对象,从影响RAPD实验结果的4个因素(引物、dNTP、Taq酶、DNA)入手,进行单因素设计,从而寻找最佳的反应浓度,达到最佳的反应效果。优化后4种因素的最佳反应浓度分别为:引物100μmol/L、dNTP 2 500μmol/L、Taq酶2U、DNA稀释80倍。在优化的反应条件下,得到的产物重复性和稳定性都比较不错。

In the experiment, the stable and reliable experiment results of Hemicentrotus puleherrimus were obtained. Four major fac- tors （ primer, dNTP, Taq enzyme and DNA） that greatly affected the results of RAPD were chosen and carried out single factor design to seek optimal reaction concentration. After the optimization concentrations of single factor design, we find out that 100 μmol/L of primer, 2 500 μmol/L of dNTP, 2 U of Taq enzyme, Diluted 80-fold of DNA. Under the optimized reaction conditions, the results of RAPD were both good in stability and repeatability.