变性梯度凝胶配制简易装置的设计

Design of a simple apparatus for the preparation of denaturing gradient gel

目的报道一种变性梯度凝胶配制简易装置的制作和变性梯度的计算方法。方法根据液体虹吸原理,利用废旧管线和2只试剂瓶组合成一套将低变性浓度胶液与高变性浓度胶液连续稀释以形成变性梯度的简易装置。结果经过实际应用,证明该装置操作简便,而且可以准确计算和控制凝胶的变性梯度范围。该装置的使用效果与单链构象多态性(single strand conformation polymorphism,SSCP)方法进行对比,效果优于后者。结论该简易装置完全可以替代价格昂贵的用于DGGE分析的成套设备,可以在普通实验室推广应用。

Objective To report a simple apparatus compounded by the denaturing gradient gel and a method to calculate the denaturing gradient concentration. Methods According to the principle of liquid siphon,the waste pipeline and two reagent bottles were combined into a simple device which can serial dilute the low and high denaturing gradient gel solution to formulate denaturing gradient. Results After practical application,it was proved that the device was easy to operate, and the gradient range of the gel can be accurately calculate and controlled. The effect of this simple device was better than that of single strand conformation polymorphism （SSCP）. Conclusion The simple device can completely replace the commercial set of equipment for DGGE analysis,it can be popularized and applied in underdeveloped laboratories.