AMPLIFIED PROBES FOR DNA ARRAYS

我们已经开发了一种简单的和一般可访问的方法，允许廉价的阵列EST克隆膜上用于差异基因表达的研究。该方法基于mRNA的非化学计量取样，允许在廉价的阵列上检测极其罕见的mRNA，同时还降低了背景杂交。作为我们的第一个目标，我们提出了技术发展，以进一步探索非化学计量采样和减少探针的复杂性。这些努力将致力于增加每个探针检测到的基因数量，并使采样偏向特定的基因组。该方法使用的RNA比目前其他非扩增方法少两个数量级。我们的目标是进一步降低RNA的要求，以便我们能够将该方法应用于非常小的组织样本。作为最后的目标，我们提出了一个简单的示范项目，这项技术。我们将用20种药物和激素处理一个细胞系，以确定超过30亿种潜在的共调节模式（向上、向下、不变），其中只有一小部分实际上被基因占据。基因解析成受管制的监管组，定义的数据库的任意处理，可能会共享生理相关的监管组件。

We have developed a simple and generally accessible method to allow inexpensive arrays of EST clones on membranes to be used for differential gene expression studies. This method, based non- stoichiometric sampling of mRNAs, allows extremely rare mRNAs to be detected on inexpensive arrays while also lowering the background hybridization. As our first aims, we propose technology development to further explore non-stoichiometric sampling and reduction of probe complexity. These efforts will be directed towards increasing the number of genes detected by each probe and biasing sampling towards particular gene sets. The method uses two orders of magnitude less RNA than other current non-amplification methods. We aim to reduced the RNA requirement even further so that we may be able to apply the method to very small tissue samples. As a final aim we propose a simple demonstration project for this technology. We will treat a cell line with twenty drugs and hormones to define over three billion potential co-regulatory patterns (up, down, unchanged), of which only a small subset will actually be occupied by genes. Genes that parse into regulated regulatory groups, as defined by the database of arbitrary treatments, are likely to share physiologically relevant regulatory components.