Grothues D, Cantor C R, Smith C L
Department of Molecular and Cell Biology, University of California, Berkeley 94720.
Proc Natl Acad Sci U S A. 1994 May 10;91(10):4461-5. doi: 10.1073/pnas.91.10.4461.
A very rapid and efficient method for sorting and ordering large numbers of clones is presented. This top-down mapping approach divides the entire ordering problem into many smaller tasks and analyzes in parallel a gridded membrane array of clones by hybridization with probe pools. The strategy was tested on a 15-fold-coverage Schizosaccharomyces pombe cosmid library. About 1600 clones were assigned to chromosomes and to regions defined by the Not I and Sfi I restriction maps. Then, the clones were ordered into 20 contigs, which is consistent with statistical expectations for the degree of genome coverage used. The parallel ordering of clones and the computer-based analysis of digitized images make this approach very efficient; it is about 8-fold faster than existing methods. Only 61 hybridizations were needed to order 1600 clones.
本文介绍了一种用于对大量克隆进行分类和排序的非常快速且高效的方法。这种自上而下的映射方法将整个排序问题分解为许多较小的任务，并通过与探针池杂交并行分析克隆的网格膜阵列。该策略在一个覆盖度为15倍的粟酒裂殖酵母黏粒文库上进行了测试。大约1600个克隆被定位到染色体以及由Not I和Sfi I限制酶切图谱定义的区域。然后，这些克隆被排列成20个重叠群，这与所使用的基因组覆盖度的统计预期一致。克隆的并行排序以及基于计算机的数字化图像分析使得这种方法非常高效；它比现有方法快约8倍。对1600个克隆进行排序仅需要61次杂交。
