Solid phase extraction of DNA from biological samples in a post-based, high surface area poly(methyl methacrylate) (PMMA) microdevice.
Solid phase extraction of DNA from biological samples in a post-based, high surface area poly(methyl methacrylate) (PMMA) microdevice.
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DOI：
10.1039/c0lc00597e
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发表时间：
2011-05-07
期刊：
影响因子：
6.1
通讯作者：
Landers JP
中科院分区：
文献类型：
--
作者：
Reedy CR;Price CW;Sniegowski J;Ferrance JP;Begley M;Landers JP
本工作描述了用壳聚糖功能化嵌入微柱的聚甲基丙烯酸甲酯（PMMA）微流控DNA纯化装置的性能。聚甲基丙烯酸甲酯（PMMA）是一种有吸引力的衬底，用于创建用于DNA捕获的高表面积（SA）柱，因为x射线光刻技术可以用于极高可重复性的高SA结构的制造。然而，当试图结合创建一个封闭系统时，这种优势被柱子的微妙性质所抵消，并且通过调用DNA结合的基团使PMMA表面功能化的挑战。描述了用壳聚糖以ph依赖的方式结合DNA的桩表面的共价功能化方法，以及避免破坏底层桩结构的键合方法。探索了许多几何桩设计，目的是确定提供必要表面积的桩结构，而不会同时增加流体阻力，从而导致设备故障。最初的原理证明是通过恢复预纯化的人类基因组DNA （hgDNA）来证明的，通过纯化全血中的hgDNA并证明其可pcr扩增来说明现实世界的实用性。
This work describes the performance of poly(methyl methacrylate) (PMMA) microfluidic DNA purification devices with embedded microfabricated posts, functionalized with chitosan. PMMA is attractive as a substrate for creating high surface area (SA) posts for DNA capture because X-ray lithography can be exploited for extremely reproducible fabrication of high SA structures. However, this advantage is offset by the delicate nature of the posts when attempting bonding to create a closed system, and by the challenge of functionalizing the PMMA surface with a group that invokes DNA binding. Methods are described for covalent functionalization of the post surfaces with chitosan that binds DNA in a pH-dependent manner, as well as for bonding methods that avoid damaging the underlying post structure. A number of geometric posts designs are explored, with the goal of identifying post structures that provide the requisite surface area without a concurrent rise in fluidic resistance that promotes device failure. Initial proof-of-principle is shown by recovery of prepurified human genomic DNA (hgDNA), with real-world utility illustrated by purifying hgDNA from whole blood and demonstrating it to be PCR-amplifiable.
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