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一种用于核酸纯化的塑料微芯片。

A plastic microchip for nucleic acid purification.

作者信息

Liu Yuxin, Cady Nathaniel C, Batt Carl A

机构信息

Department of Physics and Astronomy, Vanderbilt Institute for Integrative Biosystem Research and Education, Vanderbilt University, 6301 Stevenson Center, Nashville, TN 37235, USA.

出版信息

Biomed Microdevices. 2007 Oct;9(5):769-76. doi: 10.1007/s10544-007-9088-9.

Abstract

A microchip for purifying nucleic acids from bacterial pathogens was designed and fabricated in plastic. The fabricated plastic microchips were tested for their ability to purify nucleic acids from the bacteria Listeria monocytogenes (L. monocytogenes), Escherichia coli (E. coli), and Salmonella typhimurium (S. typhimurium). These chips were constructed using rapid and low-cost plastic fabrication techniques including hot embossing and plastic casting. Silicon molds fabricated by photolithography and dry etching were used for chip prototyping. Zeonor plastic (poly (cycloolefin) resin) and epoxy microchips were fabricated using hot embossing and plastic casting, respectively. A low temperature sputtering technique was used to coat a layer of silicon dioxide onto the channel region for nucleic acid binding in chaotropic salt solutions. The purification channels contain an array of features to increase the surface area for DNA binding and purification. DNA was quantified with PicoGreen fluorescent dye and the quality of the material as a substrate for polymerase chain reaction (PCR) was tested using target specific primers. DNA could be recovered from the microchip and detected using PCR from a minimum of 10(6) of L. monocytogenes, E. coli, and S. typhimurium cells, respectively. With the simplicity of the plastic chip's fabrication and DNA purification, our microchip makes it ideal for a miniaturized DNA testing system.

摘要

设计并制造了一种用于从细菌病原体中纯化核酸的塑料微芯片。对制造出的塑料微芯片进行测试,评估其从单核细胞增生李斯特菌、大肠杆菌和鼠伤寒沙门氏菌中纯化核酸的能力。这些芯片采用快速且低成本的塑料制造技术构建而成,包括热压印和塑料铸造。通过光刻和干法蚀刻制造的硅模具用于芯片原型制作。分别使用热压印和塑料铸造工艺制造了Zeonor塑料(聚环烯烃树脂)微芯片和环氧微芯片。采用低温溅射技术在通道区域涂覆一层二氧化硅,以便在离液盐溶液中进行核酸结合。纯化通道包含一系列特征,以增加DNA结合和纯化的表面积。使用PicoGreen荧光染料对DNA进行定量,并使用靶标特异性引物测试该材料作为聚合酶链反应(PCR)底物的质量。分别从至少10⁶个单核细胞增生李斯特菌、大肠杆菌和鼠伤寒沙门氏菌细胞中回收DNA,并使用PCR进行检测。由于塑料芯片制造和DNA纯化过程简单,我们的微芯片使其成为小型化DNA检测系统的理想选择。

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