USE OF ETHIDIUM-BROMIDE FLUORESCENCE ENHANCEMENT TO DETECT DUPLEX DNA AND DNA BACTERIOPHAGES DURING ZONE SEDIMENTATION IN SUCROSE GRADIENTS - MOLECULAR-WEIGHT OF DNA AS A FUNCTION OF SEDIMENTATION-RATE
USE OF ETHIDIUM-BROMIDE FLUORESCENCE ENHANCEMENT TO DETECT DUPLEX DNA AND DNA BACTERIOPHAGES DURING ZONE SEDIMENTATION IN SUCROSE GRADIENTS - MOLECULAR-WEIGHT OF DNA AS A FUNCTION OF SEDIMENTATION-RATE
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DOI：
10.1021/bi00600a005
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发表时间：
1978-01-01
期刊：
影响因子：
2.9
通讯作者：
GARRISON, PN
中科院分区：
文献类型：
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作者：
SERWER, P;GRAEF, PR;GARRISON, PN
通过含有溴化乙锭的5-25%蔗糖梯度沉淀双链DNA分子和DNA噬菌体。通过用UV光照射梯度并观察由DNA诱导的溴化乙锭荧光增强来确定DNA在梯度内的位置。测定了来自噬菌体T4、T5、T7和8.3%T7缺失突变体的线性双链体DNA的相对沉降速率。必要时，对DNA沉降的距离进行校正，以适应DNA穿过40%蔗糖梯度后沉降速率的逐渐降低。当沉淀0.025-0.70 μ g每种类型的DNA时，由2个DNA分子r1“和r2”沉淀的校正距离与DNA MW m1和m2相关，公式为：r1“/r2”=（m1/m2）0.38。完整的噬菌体也沉淀在溴化乙锭-蔗糖梯度和荧光增强检测。
Duplex DNA molecules and DNA bacteriophages were sedimented through 5-25% sucrose gradients containing ethidium bromide. The location of DNA within the gradients was determined by illuminating gradients with UV light and observing the ethidium bromide fluorescence enhancement induced by the DNA. The relative sedimentation rates of linear, duplex DNA from bacteriophages T4, T5, T7 and an 8.3% T7 deletion mutant were determined. The distances sedimented by DNA were corrected, when necessary, for a progressive decrease in sedimentation rate that occurs after the DNA has traversed 40% of the sucrose gradient. The corrected distances sedimented by 2 DNA molecules, r1'' and r2'', are related to the DNA MW, m1 and m2, by the equation: r1''/r2'' = (m1/m2)0.38 when 0.025-0.70 .mu.g of each type of DNA is sedimented. Intact bacteriophages were also sedimented in ethidium bromide-sucrose gradients and detected by fluorescence enhancement.