Ericsson Olle, Jarvius Jonas, Schallmeiner Edith, Howell Mathias, Nong Rachel Yuan, Reuter Hendrik, Hahn Meinhard, Stenberg Johan, Nilsson Mats, Landegren Ulf
Department of Genetics and Pathology, The Rudbeck Laboratory, Uppsala University, SE-75185 Uppsala, Sweden.
Nucleic Acids Res. 2008 May;36(8):e45. doi: 10.1093/nar/gkn106. Epub 2008 Mar 16.
DNA microarrays serve to monitor a wide range of molecular events, but emerging applications like measurements of weakly expressed genes or of proteins and their interaction patterns will require enhanced performance to improve specificity of detection and dynamic range. To further extend the utility of DNA microarray-based approaches we present a high-performance tag microarray procedure that enables probe-based analysis of as little as 100 target cDNA molecules, and with a linear dynamic range close to 10(5). Furthermore, the protocol radically decreases the risk of cross-hybridization on microarrays compared to current approaches, and it also allows for quantification by single-molecule analysis and real-time on-chip monitoring of rolling-circle amplification. We provide proof of concept for microarray-based measurement of both mRNA molecules and of proteins, converted to tag DNA sequences by padlock and proximity probe ligation, respectively.
DNA微阵列用于监测广泛的分子事件，但诸如对低表达基因、蛋白质及其相互作用模式的测量等新兴应用将需要更高的性能，以提高检测的特异性和动态范围。为了进一步扩展基于DNA微阵列方法的实用性，我们提出了一种高性能标签微阵列程序，该程序能够对低至100个靶标cDNA分子进行基于探针的分析，线性动态范围接近10的5次方。此外，与目前的方法相比，该方案从根本上降低了微阵列上交叉杂交的风险，并且还允许通过单分子分析进行定量以及对滚环扩增进行实时芯片监测。我们分别通过用锁式探针和邻近探针连接将mRNA分子和蛋白质转化为标签DNA序列，为基于微阵列的测量提供了概念验证。
