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大肠杆菌DNA/RNA在微制造生物电子芯片上的制备及杂交分析

Preparation and hybridization analysis of DNA/RNA from E. coli on microfabricated bioelectronic chips.

作者信息

Cheng J, Sheldon E L, Wu L, Uribe A, Gerrue L O, Carrino J, Heller M J, O'Connell J P

机构信息

Nanogen, Inc., San Diego, CA 92121, USA.

出版信息

Nat Biotechnol. 1998 Jun;16(6):541-6. doi: 10.1038/nbt0698-541.

Abstract

Escherichia coli were separated from a mixture containing human blood cells by means of dielectrophoresis and then subjected to electronic lysis followed by proteolytic digestion on a single microfabricated bioelectronic chip. An alternating current electric field was used to direct the bacteria to 25 microlocations above individually addressable platinum microelectrodes. The platinum electrodes were 80 microns in diameter and had center-to-center spacings of 200 microns. After the isolation, the bacteria were lysed by a series of high-voltage pulses. The lysate contained a spectrum of nucleic acids including RNA, plasmid DNA, and genomic DNA. The lysate was further examined by electronically enhanced hybridization on separate bioelectronic chips. Dielectrophoretic separation of cells followed by electronic lysis and digestion on an electronically active chip may have potential as a sample preparation process for chip-based hybridization assays in an integrated DNA/RNA analysis system.

摘要

通过介电电泳从含有人类血细胞的混合物中分离出大肠杆菌,然后在单个微制造的生物电子芯片上进行电裂解,随后进行蛋白水解消化。使用交流电场将细菌引导至可单独寻址的铂微电极上方的25个微位置。铂电极直径为80微米,中心间距为200微米。分离后,通过一系列高压脉冲裂解细菌。裂解物包含一系列核酸,包括RNA、质粒DNA和基因组DNA。通过在单独的生物电子芯片上进行电子增强杂交进一步检查裂解物。在电子活性芯片上进行细胞的介电电泳分离,然后进行电裂解和消化,作为集成DNA/RNA分析系统中基于芯片的杂交测定的样品制备过程可能具有潜力。

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