分析测试学报2022，Vol.41Issue(4)：498-508,11.DOI:10.19969/j.fxcsxb.22010501
一种基于RepRap开源3D打印平台的低成本、自动化核酸提取系统
A Low-cost,Automated Nucleic Acid Extrac?tion System Converted from the Open-Source RepRap 3D Printer
摘要
Abstract
Automatic nucleic acid extraction is essential for nucleic acid detection such as DNA amplification and high-throughput sequencing. Currently,most commercial nucleic acid extractors are mainly based on magnetic separa?tion technology,adopting a highly integrated structural design or a closed solution,which is too costly and not condu?cive to use in resource-limited areas. In addition,it is difficult to update the functionality of the instruments once the functional module is determined. To address these challenges, a desktop automated nucleic acid extraction system was built in this work,which was converted from a low-cost 3D printer based on the Self-Replicating Rapid Prototyp?ing(RepRap)open-source project. With the RepRap open-source design,the module function of the system could be flexibly designed and programmable,thus allowing updates as needed,which significantly shortened the manufactur?ing and testing cycle of the system. In the developed system,the modular function of the heating,mechanical motion and multi-channel magnetic separation could be fully integrated with the open-source 3D printing hardware. an 8-chan?nel magnetic separation based nucleic acid extraction module was designed,which could be assembled onto the origi?nal 3-axis motion platform by replacing the 3D printer extruder. The motion path planning required for automatic ex?traction was realized by G-code programming. The experimental protocols and control software were developed for the automated nucleic acid extraction in this work. The 3-axis motion platform and nucleic acid extraction module could be effectively controlled by the host computer and the module drive circuit,respectively. The heating module consisted of 4 custom-made aluminum heating bases and the heating plate on the 3D printer. The heating bases were adapted to a 96 deep-well plate,so that the deep-well plate is well attached to the heating block for high heating efficiency. By us?ing the λDNA as the standard nucleic acid samples, the extraction performance of the system had been verified by evaluating extraction purity,and efficiency including consistency and stability. From the results,the system showed a better extraction purity and efficiency for the samples with high concentration than that with low concentration . Addi?tionally, the 8-channel extraction experiment for the plasmid DNA from the cultured E. coli cells had been success?fully carried out on the automated system in this study. This validation further demonstrated that the automated system can be utilized for extracting nucleic acids from real cell samples. Overall,the nucleic acid extraction system devel?oped in this study is expected to provide a cost-effective means for rapid point-of-care molecular testing in resource-limited environments outside the laboratory.关键词
分子诊断/磁分离/核酸检测/开源硬件Key words
molecular diagnosis/magnetic separation/nucleic acid detection/open-source hardware分类
化学化工引用本文复制引用
陈阳天,王遵亮..一种基于RepRap开源3D打印平台的低成本、自动化核酸提取系统[J].分析测试学报,2022,41(4):498-508,11.基金项目
Foundation item:the National Key Research and Development Program of China(2021YFE0200400) （2021YFE0200400）
