Zhang Yi, Bailey Vasudev, Puleo Christopher M, Easwaran Hariharan, Griffiths Elizabeth, Herman James G, Baylin Stephen B, Wang Tza-Huei
Department of Biomedical Engineering, Johns Hopkins University, Baltimore, MD 21218, USA.
Lab Chip. 2009 Apr 21;9(8):1059-64. doi: 10.1039/b821780g. Epub 2009 Mar 6.
We performed on-chip DNA methylation analysis using methylation-specific PCR (MSP) within an arrayed micro droplet-in-oil platform that is designed for more practical application of microfluidic droplet technologies in clinical applications. Unique features of this ready-to-use device include arrayed primers that are pre-deposited into open micro-reaction chambers and use of the oil phase as a companion fluid for both sample actuation and compartmentalization. These technical advantages allow for infusion of minute amounts of sample for arrayed MSP analysis, without the added complexities inherent in microfluidic droplet-based studies. Ease of use of this micro device is exemplified by analysis of two tumor suppressor promoters, p15 and TMS1 using an on-chip methylation assay. These results were consistent with standard MSP protocols, yet the simplicity of the droplet-in-oil microfluidic PCR platform provides an easy and efficient tool for DNA methylation analysis in a large-scale arrayed manner.
我们在一个阵列式油包微滴平台上使用甲基化特异性PCR（MSP）进行了芯片上的DNA甲基化分析，该平台旨在使微流控液滴技术在临床应用中得到更实际的应用。这种即用型设备的独特之处包括预先沉积在开放微反应室中的阵列引物，以及使用油相作为样品驱动和分隔的伴随流体。这些技术优势使得能够注入微量样品进行阵列MSP分析，而没有基于微流控液滴的研究所固有的额外复杂性。通过使用芯片上的甲基化检测分析两个肿瘤抑制启动子p15和TMS1，体现了这种微型设备的易用性。这些结果与标准MSP方案一致，然而油包微流控PCR平台的简单性为大规模阵列式DNA甲基化分析提供了一种简便而高效的工具。
