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通过电子束蒸发镀膜法介导的DNA附着于纳米晶化学气相沉积金刚石薄膜。

EDC-mediated DNA attachment to nanocrystalline CVD diamond films.

作者信息

Christiaens P, Vermeeren V, Wenmackers S, Daenen M, Haenen K, Nesládek M, vandeVen M, Ameloot M, Michiels L, Wagner P

机构信息

Hasselt University, Institute for Materials Research and Transnationale Universiteit Limburg, School for Life Sciences, Wetenschapspark 1, B-3590 Diepenbeek, Belgium.

出版信息

Biosens Bioelectron. 2006 Aug 15;22(2):170-7. doi: 10.1016/j.bios.2005.12.013. Epub 2006 Jan 24.

Abstract

Chemical vapour deposited (CVD) diamond is a very promising material for biosensor fabrication owing both to its chemical inertness and the ability to make it electrical semiconducting that allows for connection with integrated circuits. For biosensor construction, a biochemical method to immobilize nucleic acids to a diamond surface has been developed. Nanocrystalline diamond is grown using microwave plasma-enhanced chemical vapour deposition (MPECVD). After hydrogenation of the surface, 10-undecenoic acid, an omega-unsaturated fatty acid, is tethered by 254 nm photochemical attachment. This is followed by 1-ethyl-3-[3-dimethylaminopropyl]carbodiimide (EDC)-mediated attachment of amino (NH(2))-modified dsDNA. The functionality of the covalently bound dsDNA molecules is confirmed by fluorescence measurements, PCR and gel electrophoresis during 35 denaturation and rehybridisation steps. The linking method after the fatty acid attachment can easily be applied to other biomolecules like antibodies and enzymes.

摘要

化学气相沉积(CVD)金刚石是一种非常有前途的生物传感器制造材料,这既归因于其化学惰性,也归因于使其成为电半导体的能力,从而能够与集成电路连接。对于生物传感器构建,已经开发出一种将核酸固定到金刚石表面的生化方法。使用微波等离子体增强化学气相沉积(MPECVD)生长纳米晶金刚石。在表面氢化后,通过254nm光化学附着连接ω-不饱和脂肪酸10-十一碳烯酸。随后是1-乙基-3-[3-二甲基氨基丙基]碳二亚胺(EDC)介导的氨基(NH(2))修饰的双链DNA的附着。在35个变性和重新杂交步骤中,通过荧光测量、聚合酶链反应(PCR)和凝胶电泳确认共价结合的双链DNA分子的功能。脂肪酸附着后的连接方法可以很容易地应用于其他生物分子,如抗体和酶。

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