Start Codon Targeted (SCoT) Polymorphism: A Simple, Novel DNA Marker Technique for Generating Gene-Targeted Markers in Plants
Start Codon Targeted (SCoT) Polymorphism: A Simple, Novel DNA Marker Technique for Generating Gene-Targeted Markers in Plants
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DOI：
10.1007/s11105-008-0060-5
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发表时间：
2009-03-01
影响因子：
2.1
通讯作者：
Mackill, David J.
中科院分区：
文献类型：
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作者：
Collard, Bertrand C. Y.;Mackill, David J.
随机扩增多态性DNA(随机扩增多态性DNA)标记在植物分子遗传学研究中有着广泛的应用，但具有重复性差、一般不与基因区域相关等缺点。基于植物基因ATG起始密码子两侧的短保守区，开发了一种新的植物DNA标记的产生方法。该方法在单引物聚合酶链式反应(PCR)中使用单一的18个引物，温度为50A℃，用标准琼脂糖凝胶电泳法分离扩增产物。这一方法在水稻中得到了验证，使用了一组遗传多样性的基因类型和一个回交群体。采用重复样品和不同时间进行聚合酶链式反应的方法评价方法的重复性。起始密码子靶向(SCOT)标记通常是可重复性的，但例外情况表明，引物长度和退火温度不是决定重复性的唯一因素。SCOT标记的聚合酶链式反应扩增图谱显示出与随机扩增标记类似的显性标记。我们建议，这种方法可以与这些标记一起用于遗传分析、批量分离分析和数量性状基因座定位，特别是在偏爱琼脂糖凝胶电泳法的实验室。
Random amplified polymorphic DNA (RAPD) markers have been used for numerous applications in plant molecular genetics research despite having disadvantages of poor reproducibility and not generally being associated with gene regions. A novel method for generating plant DNA markers was developed based on the short conserved region flanking the ATG start codon in plant genes. This method uses single 18-mer primers in single primer polymerase chain reaction (PCR) and an annealing temperature of 50A degrees C. PCR amplicons are resolved using standard agarose gel electrophoresis. This method was validated in rice using a genetically diverse set of genotypes and a backcross population. Reproducibility was evaluated by using duplicate samples and conducting PCR on different days. Start codon targeted (SCoT) markers were generally reproducible but exceptions indicated that primer length and annealing temperature are not the sole factors determining reproducibility. SCoT marker PCR amplification profiles indicated dominant marker like RAPD markers. We propose that this method could be used in conjunction with these markers for applications such as genetic analysis, bulked segregant analysis, and quantitative trait loci mapping, especially in laboratories with a preference for agarose gel electrophoresis.