自动化PCR体系配置及PCR产物纯化系统的初步构建

Initial establishment of automated PCR system configuration and PCR product purification system

目的建立适合于微生物实验室以及分子分型实验室的高通量的自动化PCR体系配置与PCR产物纯化系统。方法使用Janus自动液体工作站和一次性机械手枪头进行PCR体系的配置以及PCR产物的纯化,比较气动活塞移动速度、枪头与工作液面之间的距离以及是否使用一次性枪头等不同条件下,PCR体系构建中的交叉污染率以及工作效率。结果气动活塞移动速度、机械手枪头与液面的距离与交叉污染率密切相关,即气动活塞移动速度越快、枪头与液面之间的距离越大,污染率越高;与传统方法相比,在使用一次性机械手枪头的情况下,建立的高通量自动化PCR体系的工作效率提高2.5倍。结论本实验建立的工作体系能够提高工作效率和准确性,在微生物实验室和分子分型实验室中有较好的应用前景。

Objective To establish high throughput automated PCR system configuration and PCR product purification system suitable for both molecular and microbiology laboratory. Methods Use Janus automated workstation and disposal robotic tips to establish automated PCR system configuration and PCR product purification system. We compared the cross-contamination and efficiency under different parameters,such as plunger speed,the distance between tips and liquid surface and whether disposal robotic tips were used. Results Plunger speed and the distance between tips and liquid surface are two major factors that cause cross-contamination. The higher was the plunger speed and the larger was the distance between the tips and the liquid surface,the higher was cross-contamination. Compared to the traditional manual method,the automated system improved the efficiency by 2. 5 fold with the use of disposal robotic tips. Conclusion The system can improve the efficiency and the accuracy which is useful in both microbiology and molecular typing laboratory.