NEW METHOD FOR GENERATING OVERLAPPING DNA SUBCLONES

我们描述了一种生成重叠片段的新方法 用于DNA测序。此过程是独一无二的，因为没有 需要核酸外切酶或核酸内切酶。这种方法 在几个方面改进了其他程序。无计时 酶反应是产生缺失所必需的。这个 删除通过蓝/白筛选来识别。准备工作 大量的DNA是不必要的。此外，我们还描述了 一种新型DNA载体的构建 分离DNA片段中有缺失的克隆将接近于 100% DNA测序的改进具有重要的商业意义 申请。基因测序将会更快。大号- 规模排序项目将变得更具成本效益， 特别是当我们的矢量与我们的方法集成在一起时。一个 生产嵌套集合的简单方法也可能有助于 将其本身转化为自动化。该项目将演示这一新的 方法，并表明我们的方法结合在一起， 有了我们的新载体，提高了筛查效率 删除。

We describe a new method for generating overlapping fragments for use in DNA sequencing. This procedure is unique because no exonucleases or endonucleases are required. This method improves upon other procedures in several ways. No timed enzymatic reaction is necessary to produce the deletions. The deletions are identified by blue/white screening. The preparation of large amounts of DNA is unnecessary. In addition we describe the constrution of a new DNA vector in which the efficiency of isolating clones with deletions in a DNA fragment will approach 100%. Improvement of DNA sequencing has important commercial applications. Sequencing of genes would be more rapid. Large- scale sequencing projects would become more cost effective, especially when our vector is integrated with our method. A simple method for the production of nested sets might also lend itself to automation. This project will demonstrate this new method on a test gene and show that our method, in combination with our new vector, increases the efficiency of screening for deletions.