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基于热阻梯度加热系统的温度梯度芯片毛细管电泳用于DNA突变检测
引用本文:徐章润,李娜,张惠丹,樊晓峰,方瑾.基于热阻梯度加热系统的温度梯度芯片毛细管电泳用于DNA突变检测[J].高等学校化学学报,2010,31(4):667-671.
作者姓名:徐章润  李娜  张惠丹  樊晓峰  方瑾
作者单位:1. 东北大学分析科学研究中心,沈阳,110004
2. 中国医科大学卫生部细胞生物学重点实验室医学细胞生物学教育部重点实验室,细胞生物学教研室,沈阳,110001
基金项目:国家"九七三"计划项目,国家自然科学基金,辽宁省教育厅重点实验室支持计划项目 
摘    要:建立了一种简单、可靠的空间温度梯度芯片毛细管电泳DNA突变分析系统,制作了热阻呈梯度均匀变化的硅橡胶(PDMS)基片,利用其热阻变化对热传导的影响,在基片表面形成稳定的空间温度梯度.通过改变PDMS基片的厚度差,可得到范围不同的温度梯度,且形成的温度梯度在6h内保持稳定.利用该温度梯度加热装置对玻璃微流控芯片进行加热,在10℃温度梯度范围内对209bp的DNA突变标准样品进行分离检测,单次样品分析时间为8.3min,并成功用于3例大肠癌患者石蜡组织切片中K-ras基因突变的检测.

关 键 词:微流控芯片  毛细管电泳  温度梯度  基因突变
收稿时间:2009-11-16

On-chip Temperature Gradient Capillary Electrophoresis with Thermal Resistance Gradient Heating System for DNA Mutation Detection
XU Zhang-Run,LI Na,ZHANG Hui-Dan,FAN Xiao-Feng,FANG Jin.On-chip Temperature Gradient Capillary Electrophoresis with Thermal Resistance Gradient Heating System for DNA Mutation Detection[J].Chemical Research In Chinese Universities,2010,31(4):667-671.
Authors:XU Zhang-Run  LI Na  ZHANG Hui-Dan  FAN Xiao-Feng  FANG Jin
Institution:1. Research Center for Analytical Sciences, Northeastern University, Shenyang 110004, China;
2. Department of Cell Biology, Key Lab of Cell Biology of Ministry of Public Health, Key Laboratory of Medical Cell Biology of Ministry of Education, China Medical University, Shenyang 110001, China
Abstract:A simple and reliable on-chip spatial temperature gradient capillary electrophoresis was developed for DNA mutation detection using a thermal resistance gradient heating system. Polydimethylsiloxane(PDMS) slab with thermal resistance varying in gradient was made, and continuous temperature gradient was formed on slab surface using a single thermostated copper heater block owing to heat exchange influenced by varied thermal resistance. Different temperature gradients were produced by regulating the thickness of PDMS slab, and the temperature gradients were stable and uniform within 6 h. Utilizing the heating system, 209 bp DNA mutation standard samples were separated within a wide temperature range of 10 ℃ on a glass microchip with an assay time of 8.3 min. Effectiveness of the system was demonstrated by the successful detection of K-ras gene mutations in paraffin tissue sections from 3 colorectal cancer patients.
Keywords:Microfluidic chip  Capillary electrophoresis  Temperature gradient  DNA mutation
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