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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 a high-throughput automated PCR system configuration and PCR product purification system suitable for microbial laboratories and molecular typing laboratories. Methods The configuration of PCR system and purification of PCR products were carried out using Janus automatic liquid workstation and disposable mechanical handpiece. Comparing the moving conditions of pneumatic piston, the distance between the tip and the working surface and the use of one-time tip, Cross-contamination rate and efficiency in PCR system construction. Results The moving speed of the pneumatic piston and the distance between the pistol head and the liquid surface were closely related to the cross contamination rate. That is, the faster the moving speed of the pneumatic piston and the greater the distance between the tip and the liquid surface, the higher the contamination rate. Compared with the traditional method The productivity of the established high-throughput automated PCR system was improved by 2.5 times compared to the disposable robotic head. Conclusion The working system established in this experiment can improve the working efficiency and accuracy, and has a good application prospect in the microorganism laboratory and the molecular typing laboratory.