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Reliable allele detection using SNP-based PCR primers containing Locked Nucleic Acid: application in genetic mapping

机译:使用含锁定核酸的基于SNP的PCR引物进行可靠的等位基因检测:在遗传作图中的应用

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摘要

BackgroundThe diploid, Solanum caripense, a wild relative of potato and tomato, possesses valuable resistance to potato late blight and we are interested in the genetic base of this resistance. Due to extremely low levels of genetic variation within the S. caripense genome it proved impossible to generate a dense genetic map and to assign individual Solanum chromosomes through the use of conventional chromosome-specific SSR, RFLP, AFLP, as well as gene- or locus-specific markers. The ease of detection of DNA polymorphisms depends on both frequency and form of sequence variation. The narrow genetic background of close relatives and inbreds complicates the detection of persisting, reduced polymorphism and is a challenge to the development of reliable molecular markers. Nonetheless, monomorphic DNA fragments representing not directly usable conventional markers can contain considerable variation at the level of single nucleotide polymorphisms (SNPs). This can be used for the design of allele-specific molecular markers. The reproducible detection of allele-specific markers based on SNPs has been a technical challenge.
机译:背景技术二倍体茄属茄属(Solanum caripense)是马铃薯和番茄的野生近缘种,对马铃薯晚疫病具有有价值的抗性,我们对这种抗性的遗传基础感兴趣。由于加勒比拟南芥基因组中遗传变异的水平极低,因此无法通过使用常规的染色体特异性SSR,RFLP,AFLP以及基因或基因座生成密集的遗传图谱和分配单个茄属染色体特定的标记。 DNA多态性检测的难易程度取决于序列变异的频率和形式。近亲和近交者的狭窄遗传背景使持久存在的减少的多态性的检测复杂化,并且对可靠的分子标记物的开发提出了挑战。但是,代表不能直接使用的常规标记的单态性DNA片段在单核苷酸多态性(SNP)的水平上可能包含相当大的变异。这可用于设计等位基因特异性分子标记。基于SNP的等位基因特异性标记的可重复检测是一项技术挑战。

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