AN IMPROVED METHOD FOR THE DETECTION OF DNA FRAGMENTATION

被引：86

作者：

FACCHINETTI, A ^{[1
]}

TESSAROLLO, L ^{[1
]}

MAZZOCCHI, M ^{[1
]}

KINGSTON, R ^{[1
]}

COLLAVO, D ^{[1
]}

BIASI, G ^{[1
]}

机构：

[1] UNIV PADUA,INST ONCOL,CHAIR IMMUNOL,VIA GATTAMELATA 64,I-35128 PADUA,ITALY

来源：

JOURNAL OF IMMUNOLOGICAL METHODS | 1991年 / 136卷 / 01期

关键词：

APOPTOSIS DETECTION; DNA FRAGMENTATION; SOUTHERN BLOT ANALYSIS;

D O I：

10.1016/0022-1759(91)90258-H

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

An application of the Southern blot technique is described which permits the detection of DNA fragmentation due to cell death by apoptosis. DNA fragments were isolated from cell suspensions and tissues, separated on agarose gel, transferred by Southern blot and hybridized with a radiolabeled total cellular DNA probe. The application of this procedure to thymus cell samples, revealed the distinct ladder pattern of DNA fragments in multiples of about 180-200 base pairs, a characteristic feature of DNA fragmentation. In comparison to conventional DNA visualization with ethidium bromide staining, the radiolabeled probe improved the detection of DNA fragments at least eight-fold. This method detects low levels of DNA fragments, as well as physiological tissue DNA fragmentation, while avoiding cell damage due to DNA radiolabeling.

引用

页码：125 / 131

页数：7

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