Measurement of telomeric DNA content in human tissues
Measurement of telomeric DNA content in human tissues
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DOI：
10.2144/97233st05
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发表时间：
1997-09-01
期刊：
影响因子：
2.7
通讯作者：
Griffith, JK
中科院分区：
文献类型：
--
作者：
Bryant, JE;Hutchings, KG;Griffith, JK
端粒是真核染色体末端的核蛋白复合物，在体细胞中长度为 10-12 kbp，但在快速生长的癌细胞中长度仅为 1-2 kbp。 Southern blot分析是目前测量端粒长度的标准方法。然而，当 DNA 断裂或缺乏时，就不可能进行准确的测定。为了避免这些问题，开发了一种狭缝印迹测定法，可以定量端粒 DNA 的相对含量而不是长度。通过该槽印迹测定测定的端粒DNA的相对含量与通过Southern印迹分析平行测定的端粒DNA的相对长度成正比。可以在总 DNA 含量低至 15 ng 的样品中测量相对端粒 DNA 含量。相对端粒 DNA 含量（而非长度）也不受 DNA 断裂成长度为 1 kbp 或更短的片段的影响。
Telomeres, nucleoprotein complexes at the ends of eukaryotic chromosomes, are 10-12 kbp in length in somatic cells, but as small as 1-2 kbp in rapidly growing cancer cells. Southern blot analysis is currently the standard method for the measurement of telomere length. However, accurate determinations are not possible when DNA is broken or scant. To avoid these problems, a slot blot assay that quantitates the relative content, instead of length, of telomere DNA was developed. The relative contents of telomere DNA determined by this slot blot assay were directly proportional to the relative lengths of telomere DNA determined in parallel by Southern blot analysis. Relative telomere DNA content could be measured in samples containing as little as 15 ng of total DNA. Relative telomere DNA content, but not length, also was unaffected by breakage of DNA into fragments 1 kbp or less in length.