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Genome-scale design of PCR primers and long oligomers for DNA microarrays

机译:DNA芯片PCR引物和长寡聚体的基因组规模设计

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摘要

During the last years, the demand for custom-made cDNA chips/arrays as well as whole genome chips is increasing rapidly. The efficient selection of gene-specific primers/oligomers is of the utmost importance for the successful production of such chips. We developed GenomePRIDE, a highly flexible and scalable software for designing primers/oligomers for large-scale projects. The program is able to generate either long oligomers (40–70 bases), or PCR primers for the amplification of gene-specific DNA fragments of user-defined length. Additionally, primers can be designed in-frame in order to facilitate large-scale cloning into expression vectors. Furthermore, GenomePRIDE can be adapted to specific applications such as the generation of genomic amplicon arrays or the design of fragments specific for alternative splice isoforms. We tested the performance of GenomePRIDE on the entire genomes of Listeria monocytogenes (1584 gene-specific PCRs, 48 long oligomers) as well as of eukaryotes such as Schizosaccharomyces pombe (5006 gene-specific PCRs), and Drosophila melanogaster (21 306 gene-specific PCRs). With its computing speed of 1000 primer pairs per hour and a PCR amplification success of 99%, GenomePRIDE represents an extremely cost- and time-effective program.
机译:在过去的几年中,对定制的cDNA芯片/阵列以及整个基因组芯片的需求正在迅速增长。有效选择基因特异性引物/寡聚体对于成功生产此类芯片至关重要。我们开发了GenomePRIDE,这是一种高度灵活且可扩展的软件,用于设计大型项目的引物/寡聚物。该程序能够生成长寡聚体(40-70个碱基)或PCR引物,用于扩增用户定义长度的基因特异性DNA片段。另外,可以在读框内设计引物,以便于大规模克隆到表达载体中。此外,GenomePRIDE可以适应特定的应用,例如基因组扩增子阵列的产生或替代剪接同工型特异性片段的设计。我们测试了GenomePRIDE在单核细胞增生李斯特菌(1584个基因特异性PCR,48个长寡聚体)以及真核生物如粟酒裂殖酵母(5006个基因特异性PCR)和果蝇果蝇(21306个基因特异性)的整个基因组上的性能PCR)。 GenomePRIDE的计算速度为每小时1000对引物,PCR扩增成功率为99%,代表了一项极具成本效益和时间效益的程序。

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