Measurement of telomere DNA content by dot blot analysis
Measurement of telomere DNA content by dot blot analysis
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DOI：
10.1093/nar/gkr235
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发表时间：
2011-07-01
影响因子：
14.9
通讯作者：
Aviv, Abraham
中科院分区：
文献类型：
--
作者：
Kimura, Masayuki;Aviv, Abraham
端粒在人类癌症、心血管衰老和可能的长寿中起着核心作用。然而，目前测量端粒长度的方法充满了限制其使用的缺点。在这里，我们描述了一种新的方法来测量相对端粒DNA含量的斑点印迹分析。在每个点中，通过DNA染色剂（Dx）测量DNA含量，并且用端粒探针（T）测量端粒DNA含量。对每个点的Dx归一化的T（T/Dx）是端粒含量的量度。该方法每次测定需要20 ng DNA。此外，T/Dx数据与末端限制性片段Southern印迹法测得的平均端粒长度高度线性相关（r > 0.96，P < 0.0001）。该方法使用简单，具有相对较低的批间变异系数（< 6%），在中度降解的DNA中保持其精确度，并可用于高通量分析。该方法可能有助于研究人员和临床医生了解人类疾病的风险和程度。
Telomeres play a central role in human cancer, cardiovascular aging and possibly longevity. However, present methods to measure telomere length are fraught with shortcomings that limit their use. Here, we describe a novel method to measure the relative telomere DNA content by dot blot analysis. In each dot, the DNA content is measured by a DNA stain (Dx) and the telomeric DNA content is measured with a telomeric probe (T). The T normalized for Dx (T/Dx) of each dot is a measure of telomere content. The method requires similar to 20 ng of DNA per assay. Moreover, the T/Dx data are highly correlated linearly with mean telomere lengths derived from Southern blots of the terminal restriction fragments (r > 0.96, P < 0.0001). The method is also simple to use, has a relatively low interassay coefficient of variation (< 6%), retains its precision in moderately degraded DNA and can be forged for high throughput analysis. The method might help researchers and clinicians alike in understanding risks for and extent of human diseases.