Herold Keith E, Rasooly Avraham
Department of Mechanical Engineering, University of Maryland, College Park, MD 20742.
Biotechniques. 2003 Dec;35(6):1216-21. doi: 10.2144/03356bc02.
Oligonucleotide microarrays have demonstrated potential for the analysis of gene expression, genotyping, and mutational analysis. Our work focuses primarily on the detection and identification of bacteria based on known short sequences of DNA. Oligo Design, the software described here, automates several design aspects that enable the improved selection of oligonucleotides for use with microarrays for these applications. Two major features of the program are: (i) a tiling algorithm for the design of short overlapping temperature-matched oligonucleotides of variable length, which are useful for the analysis of single nucleotide polymorphisms and (ii) a set of tools for the analysis of multiple alignments of gene families and related short DNA sequences, which allow for the identification of conserved DNA sequences for PCR primer selection and variable DNA sequences for the selection of unique probes for identification. Note that the program does not address the full genome perspective but, instead, is focused on the genetic analysis of short segments of DNA. The program is Internet-enabled and includes a built-in browser and the automated ability to download sequences from GenBank by specifying the GI number. The program also includes several utilities, including audio recital of a DNA sequence (useful for verifying sequences against a written document), a random sequence generator that provides insight into the relationship between melting temperature and GC content, and a PCR calculator.
寡核苷酸微阵列已显示出在基因表达分析、基因分型和突变分析方面的潜力。我们的工作主要集中在基于已知的短DNA序列来检测和鉴定细菌。本文所述的软件Oligo Design能自动完成几个设计方面的工作,从而改进用于这些应用的微阵列寡核苷酸的选择。该程序的两个主要特点是:(i)一种平铺算法,用于设计可变长度的短重叠温度匹配寡核苷酸,这对于单核苷酸多态性分析很有用;(ii)一组用于分析基因家族和相关短DNA序列多重比对的工具,可用于鉴定保守DNA序列以选择PCR引物,以及鉴定可变DNA序列以选择用于识别的独特探针。请注意,该程序并不着眼于全基因组视角,而是专注于DNA短片段的遗传分析。该程序支持联网,包括一个内置浏览器以及通过指定GI编号从GenBank自动下载序列的功能。该程序还包括几个实用工具,包括DNA序列的语音朗读(有助于对照书面文档验证序列)、一个随机序列生成器(可深入了解解链温度与GC含量之间的关系)以及一个PCR计算器。