Mizukami T, Chang W I, Garkavtsev I, Kaplan N, Lombardi D, Matsumoto T, Niwa O, Kounosu A, Yanagida M, Marr T G
Howard Hughes Medical Institute, Cold Spring Harbor, New York 11724.
Cell. 1993 Apr 9;73(1):121-32. doi: 10.1016/0092-8674(93)90165-m.
We present the application of a nonrandom sequence-tagged site (STS) content detection method in mapping an entire genome, that of fission yeast. The novelty of our strategy is in the use of STS probes made from both ends of cosmid clones, selected on the basis of "sample without replacement" (only library clones that show no previous positive hybridization are selected and made into probes). We developed powerful techniques, based on consistency analysis, for error detection and contig assembly. In addition, we probed our library with genetically mapped markers and Notl or Sfil linking clones, thereby anchoring contigs onto chromosomes. Our map contains more than 1000 sites, including genes (most were previously unmapped), occurrences of known repetitive elements, and Notl-Sfil restriction sites.
我们展示了一种非随机序列标签位点（STS）含量检测方法在绘制整个基因组（即裂殖酵母基因组）图谱中的应用。我们策略的新颖之处在于使用了由黏粒克隆两端制备的STS探针，这些探针是基于“无放回抽样”选择的（仅选择那些先前未显示阳性杂交的文库克隆并制成探针）。我们基于一致性分析开发了强大的技术，用于错误检测和重叠群组装。此外，我们用遗传定位的标记以及Notl或Sfil连接克隆对文库进行探测，从而将重叠群锚定到染色体上。我们的图谱包含1000多个位点，包括基因（大多数先前未定位）、已知重复元件的出现情况以及Notl - Sfil限制位点。
