Two-dimensional, computer-controlled film scanner: quantitation of fluorescence from ethidium bromide-stained DNA gels.

Two-dimensional, computer-controlled film scanner: quantitation of fluorescence from ethidium bromide-stained DNA gels.
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二维计算机控制胶片扫描仪:对溴化乙锭染色的 DNA 凝胶的荧光进行定量。

DOI:
10.1016/0003-2697(84)90023-x
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发表时间:
1984
影响因子:
2.9
通讯作者:
G. Ciarrocchi
G. Ciarrocchi
中科院分区:
生物学4区
文献类型:
--
作者:
J. Sutherland;D. Monteleone;J. Trunk;G. Ciarrocchi

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介绍了一种基于数字绘图仪的二维扫描仪。该装置用于分析溴化乙啶染色dna琼脂糖凝胶的照相底片。扫描由光度数据传输到计算机进行处理、存储、显示和分析,如波段下区域的整合和确定多分散样品的平均迁移距离。用于读取光学“条形码”的集成光源和检测器模块安装在绘图仪的笔上。空间分辨率和再现性分别约为0.2和0.005 mm。通过对光源强度进行正弦调制,并通过锁相放大器对探测器发出的信号进行同步、相敏检测,光度精度达到千分之一。传感器组件的任何部分都不能接触到负极表面。与密度计相比,计算机将光度数据转换成与原始凝胶上给定点的DNA数量成正比的值。在负的两个维度上移动传感器的能力允许在一条车道的宽度上正确地集成,允许DNA的不均匀分布。
A two-dimensional scanner based on a digital plotter is described. The device is used to analyze photographic negatives of ethidium bromide-stained DNA-agarose gels. Scanning is controlled by and photometric data transferred to a computer for processing, storage, display, and analysis such as integration of the areas under bands and determination of the mean distances of migration of polydisperse samples. An integral light source and detector module designed for reading optical “bar-codes” is mounted in place of the pen of the plotter. Spatial resolution and reproducibility are about 0.2 and 0.005 mm, respectively. Photometric precision as good as one part per thousand is achieved by sinusoidal modulation of the intensity of the light source and synchronous, phase-sensitive detection of the signal from the detector by a lock-in amplifier. No part of the sensor assembly touches the surface of the negative. In contrast to a densitometer, the computer transforms photometric data to values directly proportional to the amount of DNA at given points on the original gel. The ability to move the sensor in two dimensions over the negative allows for the integration across the width of a lane correctly allowing for the nonuniform distribution of the DNA.
通过碱性琼脂糖凝胶电泳定量非放射性脱氧核糖核酸的嘧啶二聚体含量。
DOI: 10.1021/bi00273a006
发表时间: 1983
期刊: Biochemistry
影响因子: 2.9
作者:
Sutherland,BM;Shih,AG
通讯作者: Shih,AG