ASSAY FOR NANOGRAM QUANTITIES OF DNA IN CELLULAR HOMOGENATES
ASSAY FOR NANOGRAM QUANTITIES OF DNA IN CELLULAR HOMOGENATES
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DOI：
10.1016/0003-2697(79)90690-0
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发表时间：
1979-01-01
影响因子：
2.9
通讯作者：
JAMES, TW
中科院分区：
文献类型：
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作者：
BRUNK, CF;JONES, KC;JAMES, TW
使用与 DNA 复合的 4'',6-二脒基-2-苯基吲哚 (DAPI) 或双苯并咪唑 (Hoechst H 33258) 的荧光增强功能，可以在 ngram 范围内准确测量粗制细胞匀浆的 DNA 浓度。设计了一种简单的测定法，包括内标，它可以进行可靠的测量并补偿由于细胞成分或缓冲液引起的任何猝灭。荧光增强对DNA具有高度特异性；没有其他细胞成分产生显着的荧光。响应在广泛的动态范围内呈线性，使得未知 DNA 浓度的测量变得方便。 [该方法适用于嗜热四膜虫和大鼠肌肉L6细胞的匀浆]。
The DNA concentration of a crude cellular homogenate can be measured accurately in the ngram range using the fluorescence enhancement of 4'',6-diamidino-2-phenylindole (DAPI) or bisbenzimidazole (Hoechst H 33258) complexed with DNA. A simple assay was devised including an internal standard, which allows reliable measurement and compensates for any quenching due to cellular components or buffer. The fluorescence enhancement is highly specific for DNA; no other cell component produces significant fluorescence. The response is linear over a broad dynamic range making the measurement of unknown DNA concentrations convenient. [The method was applied to homogenates of Tetrahymena thermophila and rat muscle L6 cells.].