AMPLIFIED PROBES FOR DNA ARRAYS

我们已经开发了一种简单且易于使用的方法，允许廉价的EST克隆阵列在膜上用于差异基因表达研究。这种方法，基于非化学计量采样的mrna，允许极其罕见的mrna检测廉价的阵列，同时也降低了背景杂交。作为我们的第一个目标，我们提出了进一步探索非化学计量采样和降低探针复杂性的技术发展。这些努力的方向将是增加每个探针检测到的基因数量，并将抽样偏向于特定的基因集。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减少对RNA的需求，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，该技术。我们将用20种药物和激素来治疗细胞系，以确定超过30亿种潜在的共同调节模式（向上，向下，不变），其中只有一小部分实际上会被基因占据。根据任意治疗数据库的定义，解析成受调控组的基因很可能共享生理学上相关的调控成分。

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.