Goold R D, diSibio G L, Xu H, Lang D B, Dadgar J, Magrane G G, Dugaiczyk A, Smith K A, Cox D R, Masters S B
Department of Physiology, University of California San Francisco 94143-0925.
Hum Mol Genet. 1993 Aug;2(8):1271-88. doi: 10.1093/hmg/2.8.1271.
As part of our efforts to construct a high-resolution physical map of human chromosome 4, we developed a systematic approach for efficiently generating large numbers of chromosome-specific sequence-tagged sites (STSs). In this paper, we describe how rate-limiting steps in our STS development were identified and overcome, and detail our current development strategy. We present information for 822 new human chromosome 4-specific STSs, including PCR amplification conditions and subchromosomal localization data, obtained by analysis of the STS with somatic cell hybrids containing different portions of human chromosome 4. Although most STSs presented here were developed from anonymous clones whose sequences were determined in this laboratory, several STSs were developed for genes and other DNA sequences that were previously mapped to chromosome 4. Our data indicate that the availability of DNA sequence for an STS locus, in addition to the sequences of the two PCR oligonucleotides, significantly increases the transfer of that STS by allowing investigators to select new oligonucleotides best suited to the standard conditions used in their laboratories.
作为构建人类4号染色体高分辨率物理图谱工作的一部分,我们开发了一种系统方法,用于高效生成大量染色体特异性序列标签位点(STS)。在本文中,我们描述了如何识别和克服STS开发中的限速步骤,并详述了我们当前的开发策略。我们展示了822个新的人类4号染色体特异性STS的信息,包括PCR扩增条件和亚染色体定位数据,这些数据是通过用含有人类4号染色体不同部分的体细胞杂种分析STS获得的。尽管这里展示的大多数STS是从本实验室测定了序列的匿名克隆中开发出来的,但也有几个STS是针对先前已定位到4号染色体的基因和其他DNA序列开发的。我们的数据表明,除了两个PCR寡核苷酸的序列外,STS位点的DNA序列可用性通过允许研究人员选择最适合其实验室使用的标准条件的新寡核苷酸,显著提高了该STS的转移性。