A miniaturized spatial temperature gradient capillary electrophoresis system with radiative heating and automated sample introduction for DNA mutation detection
A miniaturized spatial temperature gradient capillary electrophoresis system with radiative heating and automated sample introduction for DNA mutation detection
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DOI：
10.1002/elps.201000205
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发表时间：
2010-09
期刊：
影响因子：
2.9
通讯作者：
Zhangrun Xu;Qi Li;Xiao-feng Fan;Huidan Zhang;Jin Fang
中科院分区：
文献类型：
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作者：
Zhangrun Xu;Qi Li;Xiao-feng Fan;Huidan Zhang;Jin Fang
建立了一种用于DNA突变检测的小型化自动进样空间温度梯度毛细管电泳系统。通过将自动化缝瓶阵列进样装置与空间温度梯度CE系统相结合，实现了连续电动进样。采用单块石墨加热器辐射加热系统产生温度梯度，并对温度梯度的稳定性进行了研究。6 h内各测点温度变化的RSD（n=7）为0.12-0.21%。使用14 cm Teflon AF涂层的二氧化硅毛细管作为分离通道和荧光信号的液芯波导管。在54.8至59.5°C的温度梯度下，低范围对照突变标准品（209 bp）在4分钟内分离，仅消耗5.6 nL样品。实现了自动连续进样和换样，残留率为3.3%。通过检测两名结直肠癌患者石蜡组织切片中的K-ras基因突变，进一步证明了该系统的实用性。
A miniaturized spatial temperature gradient CE system with automated sample introduction for DNA mutation detection was established. Continuous electrokinetic sample injection was achieved by combining an automated slotted‐vial array sample introduction device to the spatial temperature gradient CE system. The temperature gradient was produced by a radiative heating system with a single graphite block heater, and the stability of the temperature gradient was investigated. The temperature variation of each measure point was 0.12–0.21% RSD (n=7) within 6 h. A 14‐cm Teflon AF‐coated silica capillary was used both as the separation channel and as the liquid‐core waveguide tube of fluorescence signal. Under a temperature gradient from 54.8 to 59.5°C, a low range control mutation standard (209 bp) was separated within 4 min with only 5.6 nL sample consumption. Automated continuous sample introducing and changing were realized with a carryover of 3.3%. Utility of the system was further demonstrated by detecting K‐ras gene mutations in paraffin tissue sections from two colorectal cancer patients.