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通过光刻技术将 DNA 特异性固定在玻璃微通道中。

Site-specific immobilization of DNA in glass microchannels via photolithography.

机构信息

Laboratory of Organic Chemistry, Wageningen University, Dreijenplein 8, 6703 HB Wageningen, The Netherlands.

出版信息

Langmuir. 2009 Dec 15;25(24):13952-8. doi: 10.1021/la901558n.

Abstract

For the first time, a microchannel was photochemically patterned with a functional linker. This simple method was developed for the site-specific attachment of DNA via this linker onto silicon oxide surfaces (e.g., fused silica and borosilicate glass), both onto a flat surface and onto the inside of a fused silica microchannel. Sharp boundaries in the micrometer range between modified and unmodified zones were demonstrated by the attachment of fluorescently labeled DNA oligomers. Studies of repeated hybridization-dehybridization cycles revealed selective and reversible binding of cDNA strands at the explicit locations. On average, approximately 7 x 10(11) fluorescently labeled DNA molecules were hybridized per square centimeter. The modified surfaces were characterized with X-ray photoelectron spectroscopy, infrared microscopy, static contact angle measurements, confocal laser scanning microscopy, and fluorescence detection (to quantify the attachment of the fluorescently labeled DNA).

摘要

首次采用光化学技术在微通道上形成带有功能连接物的图案。该简单方法可用于通过此连接物将 DNA 特异性地连接到硅氧化物表面(例如,熔融二氧化硅和硼硅酸盐玻璃)上,无论是在平面上还是在熔融二氧化硅微通道内部。通过附着荧光标记的 DNA 寡聚物证明了在修饰和未修饰区域之间具有亚微米级别的锐利边界。杂交-去杂交循环的研究表明 cDNA 链在明确位置处具有选择性和可逆的结合。平均而言,每平方厘米可杂交大约 7 x 10(11)个荧光标记的 DNA 分子。使用 X 射线光电子能谱、红外显微镜、静态接触角测量、共聚焦激光扫描显微镜和荧光检测(用于定量附着的荧光标记 DNA)对修饰表面进行了表征。

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