Bio & Health Lab, Samsung Advanced Institute of Technology, Samsung Electronics Co., Ltd., San #14-1, Nongseo-dong, Giheung-gu, Yongin-si, Gyeonggi-do, Republic of Korea.
Lab Chip. 2010 Mar 7;10(5):626-33. doi: 10.1039/b916606h. Epub 2009 Dec 17.
We present a novel electrochemical cell lysis device to prepare DNA samples for lab-on-a-chip (LOC) applications. It utilizes the electrolysis of saline solution to generate hydroxide ions (OH(-)) at the cathode as alkaline lytic agents. Cathode and anode chambers are separated by a negatively-charged ion exchangeable polymer diaphragm to maintain the high pH level for efficient cell lysis in the cathode chamber, to prevent inflow of PCR-amplification inhibitors from the anode chamber, and to minimize binding of DNA molecules. Electric current flow and pH maintenance, which depended on the device design, were two important parameters of the device performance. After optimizing the design and visually confirming cell lysis of Chinese hamster ovary (CHO) cells in a very short amount of time, we directly electrolyzed four bacterial cell types suspended in saline solution. Real-time PCR (qPCR) analysis showed that our device could lyse both gram-positive and gram-negative bacterial cells with higher efficiency than other common methods and could detect DNA on the microlitre scale. Our data demonstrate several advantages of the proposed device: absence of cell lysis chemicals and heating; no adverse effects on PCR amplification; low DNA loss; low voltage and power consumption; and rapid processing. The device could potentially be applied as an on-chip DNA extraction component.
我们提出了一种新颖的电化学细胞裂解装置,用于为芯片实验室 (LOC) 应用制备 DNA 样品。它利用盐水的电解在阴极产生氢氧根离子 (OH(-)) 作为碱性裂解试剂。阴极室和阳极室由带负电荷的可交换离子聚合物隔膜隔开,以保持阴极室内的高 pH 值,实现高效的细胞裂解,防止阳极室内的 PCR 扩增抑制剂流入,并最小化 DNA 分子的结合。电流流动和 pH 值维持取决于装置设计,是装置性能的两个重要参数。在优化设计并在很短的时间内直观地确认中国仓鼠卵巢 (CHO) 细胞的裂解后,我们直接对悬浮在盐水中的四种细菌细胞进行了电解。实时 PCR (qPCR) 分析表明,与其他常见方法相比,我们的装置可以更有效地裂解革兰氏阳性和革兰氏阴性细菌细胞,并能够在微升范围内检测到 DNA。我们的数据证明了所提出的装置具有几个优势:没有细胞裂解化学物质和加热;对 PCR 扩增没有不利影响;DNA 损失低;电压和功耗低;处理速度快。该装置有可能被用作芯片上的 DNA 提取组件。