Center for Applied NanoBioscience and Medicine, The University of Arizona College of Medicine, Phoenix, AZ 85004, USA.
Electrophoresis. 2010 Oct;31(21):3510-7. doi: 10.1002/elps.201000305.
The microfluidic integration of an entire DNA analysis workflow on a fully integrated miniaturized instrument is reported using lab-on-a-chip automation to perform DNA fingerprinting compatible with CODIS standard relevant to the forensic community. The instrument aims to improve the cost, duration, and ease of use to perform a "sample-to-profile" analysis with no need for human intervention. The present publication describes the operation of the three major components of the system: the electronic control components, the microfluidic cartridge and CE microchip, and the optical excitation/detection module. Experimental details are given to characterize the level of performance, stability, reliability, accuracy, and sensitivity of the prototype system. A typical temperature profile from a PCR amplification process and an electropherogram of a commercial size standard (GeneScan 500™, Applied Biosystems) separation are shown to assess the relevance of the instrument to forensic applications. Finally, we present a profile from an automated integrated run where lysed cells from a buccal swab were introduced in the system and no further human intervention was required to complete the analysis.
本文报道了一种将整个 DNA 分析工作流程集成在一个完全集成的微型仪器上的微流控技术,该技术使用芯片实验室自动化来进行 DNA 指纹分析,与法医界相关的 CODIS 标准兼容。该仪器旨在降低成本、缩短时间,并提高易用性,实现“样本到分析结果”的无人干预分析。本出版物介绍了系统的三个主要组成部分的操作:电子控制组件、微流控盒和 CE 微芯片以及光学激发/检测模块。给出了实验细节,以表征原型系统的性能、稳定性、可靠性、准确性和灵敏度水平。展示了一个典型的 PCR 扩增过程的温度曲线和商业大小标准(GeneScan 500™,Applied Biosystems)分离的电泳图谱,以评估该仪器在法医应用中的相关性。最后,我们展示了一个自动化集成运行的结果,其中从口腔拭子中裂解的细胞被引入系统,并且无需进一步的人工干预即可完成分析。