一种基于反相微乳液的数字PCR微滴的制备方法

Preparation of droplets in digital PCR based on inverse microemulsion

目的提出一种新型的微滴生成方式——油包水(W/O)包裹技术,为数字PCR实验过程中微滴的制备提供一种更便捷的方式。方法通过配制一定比例的特定油相以及包含模板DNA等反应物的特定水相,在不同条件下形成W/O微滴,并进行PCR反应。结果所形成的微滴粒径大小均匀,并且在进行PCR反应过程中热力学性能保持不变,微滴不会因温度升降而破裂,适合于数字PCR中微滴的制备。结论生成的微滴可以有效地使模板DNA进行扩增反应。

Objective A new method for preparation of droplets is presented： wrapping technique for water-in-oil（W/O）,it provides a more convenient way for the digital PCR experiment. Methods We formulated a certain percentage of particular oil phase with water phase containing the template DNA,maked the mixture to form W/O droplets,then carried out PCR reactions. Results The size of droplets prepared was uniform. Droplets＇ thermodynamic properties remained constant during the PCR reactions,and the droplet didn＇t rupture with the temperature changes. Conclusions The electrophoresis test results show that we make the amplification reaction of the template DNA successfully.