来自溴脱氧尿苷取代细胞的DNA的热变性
Thermal denaturation of DNA from bromodeoxyuridine substituted cells.
作者信息
Cullen B R, Bick M D
出版信息
Nucleic Acids Res. 1976 Jan;3(1):49-62. doi: 10.1093/nar/3.1.49.
Abstract
The thermal denaturation of DNA from cell lines extensively substituted with bromodeoxyuridine has been examined spectrophotometrically over a wide range in ionic strength and by thermal elution from hydroxyapatite columns. BrdU substitution stabliizes DNA at all ionic strengths between 7.5 mM and 1350 mM potassium ion concentration, although a plot of log ionic strength vs Tm deviates from linearity above 150 mM. This nonlinearity is most pronounced with BrdU-substituted DNAs, resulting in a lowered delta Tm between unsubstituted and substituted DNA with increasing ionic strength. DMSO is shown to decrease the Tm of both unsubstituted and BrdU-substituted DNA equally, at a rate of .5 degrees C per 1% DMSO.
摘要
通过在广泛的离子强度范围内进行分光光度法检测，以及利用羟基磷灰石柱进行热洗脱，研究了用溴脱氧尿苷大量取代的细胞系DNA的热变性。在7.5 mM至1350 mM钾离子浓度之间的所有离子强度下，BrdU取代都会使DNA稳定，尽管在高于150 mM时，log离子强度与Tm的关系图偏离线性。这种非线性在BrdU取代的DNA中最为明显，随着离子强度增加，未取代和取代DNA之间的ΔTm降低。结果表明，DMSO以每1%DMSO 0.5℃的速率同等程度地降低未取代和BrdU取代DNA的Tm。
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