| 基于PMMA毛细管电泳芯片的多位点突变检测研究 |
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| 引用本文： | 马辰,张冠斌,邓涛,王怡瑞,邢婉丽,程京.基于PMMA毛细管电泳芯片的多位点突变检测研究[J].分析科学学报,2012,28(4):445-449. |
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| 作者姓名： | 马辰 张冠斌 邓涛 王怡瑞 邢婉丽 程京 |
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| 作者单位： | 1. 清华大学医学院生物医学工程系,北京100084;生物芯片北京国家工程研究中心,北京102206
2. 生物芯片北京国家工程研究中心,北京,102206 |
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| 基金项目： | 国家863计划重点项目 |
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| 摘 要： | 利用基于激光诱导荧光(LIF)检测的芯片毛细管电泳平台,批量制作了低成本聚甲基丙烯酸甲酯(PMMA)芯片,通过修饰管道,优化有效分离距离、分离介质等条件,可在90s内完成DNA片段的分离检测,实现单碱基分离,并在此平台上成功地对遗传性耳聋三个常见突变位点实现分型检测,为这种低成本的PMMA芯片应用于分型相关的临床诊断领域奠定了基础。
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| 关 键 词： | 芯片毛细管电泳 激光诱导荧光 遗传性耳聋 突变检测 |
Multiplex Mutation Detection by PMMA Microhip-based Capillary Electrophoresis |
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| MA Chen
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ZHANG Guan-bin
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DENG Tao
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WANG Yi-rui
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XING Wan-li
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CHENG Jing.Multiplex Mutation Detection by PMMA Microhip-based Capillary Electrophoresis[J].Journal of Analytical Science,2012,28(4):445-449. |
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| Authors: | MA Chen ZHANG Guan-bin DENG Tao WANG Yi-rui XING Wan-li CHENG Jing |
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| Institution: | 1,2(1.Department of Biomedical Engineering,School of Medicine,Tsinghua University,Beijing 10084; 2.National Engineering Research Center for Beijing Biochip Technology,Beijing 102206) |
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| Abstract: | The technology of microchip-based capillary electrophoresis(MCE) is now widely used in analytical and life science researches.In this paper,we designed a microchip-based capillary electrophoresis platform using laser-induced fluorescence(LIF) detection method and polymethyl methacrylate(PMMA) chips manufactured by UV-LIGA technology.We evaluated the resolution gained by wall coating of a PMMA chip and optimized the electrophoresis conditions including the effective separation distance and the sieving matrix.By applying the optimized conditions,we achieved a single-base resolution of DNA fragments in only 90 seconds on this platform.In addition,we genotyped clinical samples containing three common mutation sites associated with hereditary hearing loss and demonstrated the potential application of the PMMA microchip-based capillary electrophoresis system in clinical diagnostic fields. |
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| Keywords: | Microchip-based capillary electrophoresis Laser-induced fluorescence Hereditary hearing loss Mutation detection |
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