Cheng J, Shoffner M A, Hvichia G E, Kricka L J, Wilding P
Department of Pathology and Laboratory Medicine, School of Medicine, University of Pennsylvania, Philadelphia, 19104, USA.
Nucleic Acids Res. 1996 Jan 15;24(2):380-5. doi: 10.1093/nar/24.2.380.
We examined PCR in silicon dioxide-coated silicon-glass chips (12 microl in volume with a surface to volume ratio of approximately 17.5 mm(2)/microl) using two PCR reagent systems: (i) the conventional reagent system using Taq DNA polymerase; (ii) the hot-start reagent system based on a mixture of TaqStart antibody and Taq DNA polymerase. Quantitative results obtained from capillary electrophoresis for the expected amplification products showed that amplification in microchips was reproducible (between batch coefficient of variation 7.71%) and provided excellent yields. We also used the chip for PCR directly from isolated intact human lymphocytes. The amplification results were comparable with those obtained using extracted human genomic DNA. This investigation is fundamental to the integration of sample preparation, polynucleotide amplification and amplicate detection on a microchip.
我们使用两种聚合酶链式反应（PCR）试剂系统，在二氧化硅涂层的硅玻璃芯片（体积为12微升，表面积与体积比约为17.5平方毫米/微升）中检测PCR：（i）使用Taq DNA聚合酶的传统试剂系统；（ii）基于TaqStart抗体和Taq DNA聚合酶混合物的热启动试剂系统。通过毛细管电泳获得的预期扩增产物的定量结果表明，微芯片中的扩增具有可重复性（批次间变异系数为7.71%），且产量很高。我们还直接在分离出的完整人类淋巴细胞上使用该芯片进行PCR。扩增结果与使用提取的人类基因组DNA获得的结果相当。这项研究对于在微芯片上整合样品制备、多核苷酸扩增和扩增产物检测至关重要。
