A quantitative assay for fragmented DNA in apoptotic cells.
A quantitative assay for fragmented DNA in apoptotic cells.
复制标题
凋亡细胞中 DNA 片段的定量分析。
DOI：
10.1016/0003-2697(92)90518-c
复制
发表时间：
1992
影响因子：
2.9
通讯作者：
Plunkett,W
中科院分区：
文献类型：
--
作者：
Huang,P;Plunkett,W
细胞核小体间染色质的裂解是细胞凋亡过程中DNA完整性的特征性变化。我们已经开发了一种用于定量凋亡细胞中核小体间DNA片段化的测定方法。该技术包括细胞DNA的纯化、DNA末端的脱磷酸化、DNA 5′端的32 P标记、琼脂糖凝胶电泳和DNA条带中放射性的定量。该测定比通过乙锭染色的DNA条带可视化灵敏约1000至2000倍，允许检测皮克水平的DNA片段。还描述了用于定量测定片段化DNA链的数目的方法。应用这种新的检测方法来评估核小体间DNA片段化的时间过程，证明了在由抗癌核苷类似物诱导的凋亡细胞。
Cleavage of cellular chromatin at internucleosomal sites is a characteristic change of DNA integrity in cells undergoing apoptosis. We have developed an assay for quantitation of internucleosomal DNA fragmentation in apoptotic cells. This technique involves purification of cellular DNA, dephosphorylation of the DNA ends, labeling of DNA with32P at the 5′-end, gel electrophoresis through agarose, and quantitation of the radioactivity in DNA bands. This assay, which is about 1000- to 2000-fold more sensitive than visualization of DNA bands by ethidium staining, allows the detection of DNA fragments at picogram levels. A method for quantitatively determining the number of fragmented DNA strands is also described. Application of this new assay to evaluate the time course of internucleosomal DNA fragmentation was demonstrated in apoptotic cells induced by an anticancer nucleoside analogue.
影响因子：
11.2
作者：
S. Kaufmann
通讯作者：
S. Kaufmann