Robust polymeric microchannel coatings for microchip-based analysis of neat PCR products.
Robust polymeric microchannel coatings for microchip-based analysis of neat PCR products.
复制标题
坚固的聚合物微通道涂层，用于对纯 PCR 产物进行基于微芯片的分析。
DOI：
10.1021/ac001317k
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发表时间：
2001
影响因子：
7.4
通讯作者：
Landers,JP
中科院分区：
文献类型：
--
作者：
Munro,NJ;Hühmer,AF;Landers,JP
几种二氧化硅涂层已经评估了重复PCR产品分析在毛细管和电泳微芯片。二氧化硅涂层是许多电泳分离的重要组成部分，这种重要性在微芯片中被放大，其中分离距离被最小化。增加涂层对分离条件的耐受性可以提高涂层微芯片的可重复性和寿命，从而实现这些设备的全部潜力（快速分离，高通量和寿命）。在本研究中，通过实验和文献对几种包衣参数进行了评价，以生产出具有高抗性的包衣，对分离条件感兴趣，产物分离整齐。在此条件下对几种涂料进行了降解试验，并讨论了表面羟基化、表面水化、硅化溶剂、硅化试剂、催化剂、端封和聚合工艺等因素对涂层降解的影响。在测试条件下，用氯二甲基基硅烷在甲苯中与聚（乙烯基吡咯烷酮）通过抽氢法附着的聚合物层进行硅烷化制备涂层（涂层E） [Srinivasan， K.；波尔,c;Avdalovic N.Anal。Chem.1997,69, 2798−2805]抗性最强。该涂层在电泳微芯片上进行了寿命测试，并与传统的聚丙烯酰胺涂层进行了比较。两种涂层的分辨率和效率相似；然而，涂层E提供了更多的可重复迁移时间，并且在测试终止时进行了635次分析。该程序提供了一个可重复的，耐表面涂层，从而允许在微芯片上重复分析整齐的PCR产物。
Several silica coatings have been evaluated for replicate PCR product analysis in capillaries and electrophoretic microchips. Silica coatings are an essential component to many electrophoretic separations, and this importance is magnified in microchips, where separation distances are minimized. Increasing the resistance of coatings to separation conditions improves the reproducibility and longevity of the coated microchip, which allows for the full potential of these devices (rapid separations, high throughput, and longevity) to be realized. In this study, several coating parameters have been evaluated experimentally and through the literature to produce a coating with high resistance to the separation conditions of interest, neat PCR product separations. Coating degradation induced under these conditions was tested for several coatings, and the influence of surface hydroxylation, surface hydration, silanization solvent, silanizing reagent, catalysis, endcapping, and polymerization procedure are discussed. Under the testing conditions, a coating (coating E) prepared by silanization with chlorodimethyloctylsilane in toluene with a polymer layer of poly(vinylpyrrolidone) attached by a hydrogen abstraction method [Srinivasan, K.; Pohl, C.; Avdalovic, N.Anal. Chem.1997,69, 2798−2805] was most resistant. This coating was tested for longevity on electrophoretic microchips and was compared to the traditional coating of polyacrylamide. The coatings produced similar resolution and efficiencies; however, coating E provided more reproducible migration times and had performed for 635 analyses when testing was terminated. This procedure provides a reproducible, resistant surface coating, thus allowing for replicate analysis of neat PCR product on microchips.