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首页> 外文期刊>International Journal of Biological Macromolecules: Structure, Function and Interactions >ATLAS: An advanced PCR-method for routine visualization of telomere length in Saccharomyces cerevisiae
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ATLAS: An advanced PCR-method for routine visualization of telomere length in Saccharomyces cerevisiae

机译:ATLAS:一种先进的PCR方法,用于常规显示酿酒酵母中的端粒长度

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Measuring telomere length is essential in telomere biology. Southern blot hybridization is the predominant method for measuring telomere length in the genetic model Saccharomyces cerevisiae. We have further developed and refined a telomere PCR approach, which was rarely used previously (mainly in specific telomeric projects), into a robust method allowing direct visualisation of telomere length differences in routine experiments with S. cerevisiae, and showing a strong correlation of results with data obtained by Southern blot hybridization. In this expanded method denoted as ATLAS (A-dvanced T-elomere L-ength A-nalysis in S. cerevisiae), we have introduced: 1) set of new primers annealing with high specificity to telomeric regions on five different chromosomes; 2) new approach for designing reverse telomere primers that is based on the ligation of an adaptor of a fixed size to telomeric ends. ATLAS can be used at the scale of individual assays and high-throughput approaches. This simple, time/cost-effective and reproducible methodology will complement Southern blot hybridization and facilitate further progress in telomere research. (C) 2016 Elsevier B.V. All rights reserved.
机译:在端粒生物学中,测量端粒长度至关重要。 Southern印迹杂交是测量酿酒酵母基因模型中端粒长度的主要方法。我们进一步开发和完善了以前很少使用的端粒PCR方法(主要在特定的端粒项目中使用),使其成为一种健壮的方法,可以直接观察酿酒酵母常规实验中端粒的长度差异,并显示出很强的相关性通过Southern blot杂交获得的数据。在这种扩展的方法称为ATLAS(酿酒酵母中的A晚期T-elomere L长度A分析)中,我们引入了:1)对5个不同染色体上的端粒区域具有高特异性退火的新引物组; 2)设计反向端粒引物的新方法,该方法基于固定大小的衔接子与端粒末端的连接。 ATLAS可以用于单个检测和高通量方法。这种简单,时间/成本有效且可重复的方法将补充Southern blot杂交,并促进端粒研究的进一步进展。 (C)2016 Elsevier B.V.保留所有权利。

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