Palazzolo M J, Sawyer S A, Martin C H, Smoller D A, Hartl D L
Department of Genetics, Washington University School of Medicine, St. Louis, MO 63110.
Proc Natl Acad Sci U S A. 1991 Sep 15;88(18):8034-8. doi: 10.1073/pnas.88.18.8034.
The physical mapping of complex genomes is based on the construction of a genomic library and the determination of the overlaps between the inserts of the mapping clones in order to generate an ordered, cloned representation of nearly all the sequences present in the target genome. Evaluation of the relative efficiency of experimental procedures used to accomplish this goal must minimally include a comparison of the fraction of the genome covered by the ordered arrays (or "contigs"), the average size of the contigs, and the cost, in terms of time and resources, required to generate the map. Sequence-tagged-site (STS) content mapping is one strategy that has been proposed and is being utilized for this type of experiment. This paper describes three STS selection schemes and presents computer simulations of contig-building experiments based on these procedures. The results of these simulations suggest that a nonrandom STS strategy that uses paired probes requires one-third to one-fourth as many STS assays as are required in random and nonpaired approaches, and also results in a map that has both greater genome coverage and a larger average contig size. This strategy promises to reduce the time and cost required to build a high-quality physical map.
复杂基因组的物理图谱构建基于基因组文库的构建以及对作图克隆插入片段之间重叠情况的确定，以便生成目标基因组中几乎所有序列的有序、克隆化表示。对用于实现这一目标的实验方法的相对效率进行评估，至少必须包括对有序阵列（或“重叠群”）覆盖的基因组部分、重叠群的平均大小以及生成图谱所需的时间和资源成本进行比较。序列标签位点（STS）含量作图是为此类实验提出并正在使用的一种策略。本文描述了三种STS选择方案，并给出了基于这些方法的重叠群构建实验的计算机模拟。这些模拟结果表明，使用配对探针的非随机STS策略所需的STS检测数量仅为随机和非配对方法所需数量的三分之一到四分之一，并且还能得到一个具有更大基因组覆盖率和更大平均重叠群大小的图谱。该策略有望减少构建高质量物理图谱所需的时间和成本。
