Development of an Automated Platform for the Electrophoresis-based Purification of a Biomolecule by Application of Optomechatronic Techniques
Date Issued
2007
Date
2007
Author(s)
Chen, Jian-Jhou
DOI
zh-TW
Abstract
Biomolecule purification is an essential step for molecular biology experiments. It is usually a time-consuming and work-consuming process, and the yield of purification is subject to an operator’s experience. Therefore, automation that features labor-saving property and high reproducibility becomes more and more important for biotech industry.
By the application of optomechatronic techniques, this study integrates tubular gel electrophoresis, optical sensor, and automated liquid handling system to develop a real-time, automated platform for the electrophoresis-based purification of a target biomolecule.
For tubular gel electrophoresis, different electrophoresis conditions and device designs were compared and studied to achieve the best electrophoresis resolution. With optimal device design and operating conditions that minimize the sample-diffusion problem, high resolution of the tubular gel electrophoresis for both tracking dye and DNA markers was attained and could thus be integrated with an optical detection system. The optical detection system used a UV lamp as the light source, a photodiode as the detector of light absorption by biomolecules, and an amplifying circuit to process the detected optical signal. In the beginning, a tracking dye (bromophenol blue) was used to determine an appropriate electrophoresis condition for real-time optical detection. In practice, when a tracking dye migrates into the detection window, it absorbs ultraviolet light thus decreases the optical signal output of the photodiode. The experiment of results show that the optical detection system is able to detect each individual DNA marker during electrophoresis separation. Both the tubular gel electrophoresis and optical detection module are well-integrated and well-suited to develop the automated platform for electrophoresis-based purification of target biomolecules with an automated liquid handling system.
For automated tracking dye purification, the aspiration time is estimated by the peak position and mobility of the dye. The accuracy of this estimation has been verified by different experiments. It has been found that the automated purification system can provide a recovery rate of ca. 13.8 % and take 40~45 minutes to accomplish the purification.
The system developed in this thesis work is to be compatible with multi-channel electrophoresis or miniaturized device. Therefore, it can serve as a basis of high-throughput purification of samples and is thus beneficial to both acedemic workers and biotech-orientated industries.
Subjects
電泳分離:光學偵測:自動化純化
electrophoresis isolation:optical detection:automated purification
Type
thesis
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