Kastania Athina S, Tsougeni Katerina, Papadakis George, Gizeli Electra, Kokkoris George, Tserepi Angeliki, Gogolides Evangelos
Institute of Nanoscience and Nanotechnology, NCSR "Demokritos", Patriarhou Gregoriou E' & 27 Neapoleos Str., 153 41 Aghia Paraskevi, Greece.
Institute of Molecular Biology and Biotechnology-FORTH, 100 N. Plastira Str., Heraklion, 70013, Greece.
Anal Chim Acta. 2016 Oct 26;942:58-67. doi: 10.1016/j.aca.2016.09.007. Epub 2016 Sep 22.
We present a polymeric microfluidic chip capable of purifying DNA through solid phase extraction. It is designed to be used as a module of an integrated Lab-on-chip platform for pathogen detection, but it can also be used as a stand-alone device. The microfluidic channels are oxygen plasma micro-nanotextured, i.e. randomly roughened in the micro-nano scale, a process creating high surface area as well as high density of carboxyl groups (COOH). The COOH groups together with a buffer that contains polyethylene glycol (PEG), NaCl and ethanol are able to bind DNA on the microchannel surface. The chip design incorporates a mixer so that sample and buffer can be efficiently mixed on chip under continuous flow. DNA is subsequently eluted in water. The chip is able to isolate DNA with high recovery efficiency (96± 11%) in an extremely large dynamic range of prepurified Salmonella DNA as well as from Salmonella cell lysates that correspond to a range of 5 to 1.9 × 10 cells (0.263 fg to 2 × 500 ng). The chip was evaluated via absorbance measurements, polymerase chain reaction (PCR), and gel electrophoresis.
我们展示了一种能够通过固相萃取纯化DNA的聚合物微流控芯片。它被设计用作集成芯片实验室平台中用于病原体检测的一个模块，但也可以用作独立设备。微流控通道经过氧等离子体微纳纹理处理，即在微纳尺度上随机粗糙化，这一过程会产生高表面积以及高密度的羧基（COOH）。羧基与含有聚乙二醇（PEG）、氯化钠和乙醇的缓冲液一起，能够将DNA结合在微通道表面。芯片设计包含一个混合器，以便在连续流动下样品和缓冲液能在芯片上高效混合。随后DNA在水中被洗脱。该芯片能够在极宽的预纯化沙门氏菌DNA动态范围内以及对应5至1.9×10个细胞（0.263 fg至2×500 ng）范围的沙门氏菌细胞裂解物中，以高回收效率（96±11%）分离DNA。通过吸光度测量、聚合酶链反应（PCR）和凝胶电泳对该芯片进行了评估。
