A CCD-based system for the detection of DNA in electrophoresis gels by UV absorption.
A CCD-based system for the detection of DNA in electrophoresis gels by UV absorption.
复制标题
基于 CCD 的系统，用于通过紫外吸收检测电泳凝胶中的 DNA。
DOI：
10.1088/0031-9155/44/6/307
复制
发表时间：
1999
影响因子：
3.5
通讯作者：
A. Mainwood
中科院分区：
文献类型：
--
作者：
Alex R Mahon;Alex R Mahon;John H MacDonald;Robert J. Ott;A. Mainwood
提供了一种用于检测和定量琼脂糖凝胶中未标记核酸的大片段的方法和装置。该技术基于核苷酸的紫外线（UV）吸收。氘源通过光纤阵列照亮电泳凝胶的各个样品泳道。当DNA条带通过凝胶的照射区域时，由于DNA的吸收，透射的UV光的量减少。在电泳过程中，使用紫外敏感电荷耦合器件（CCD）在线检测DNA区域。由于吸收系数与DNA的质量成比例，因此该技术本质上是定量的。凝胶区域中的DNA质量与CCD图像的相应部分中的积分信号近似成比例。该系统目前的检测限小于1.25 ng，而最流行的常规技术溴化乙锭（EtBr）染色的检测限为2-10 ng。此外，DNA样品保持其天然状态。从检测程序中去除致癌染料大大降低了相关的生物危害。
A method and apparatus for the detection and quantification of large fragments of unlabelled nucleic acids in agarose gels is presented. The technique is based on ultraviolet (UV) absorption by nucleotides. A deuterium source illuminates individual sample lanes of an electrophoresis gel via an array of optical fibres. As DNA bands pass through the illuminated region of the gel the amount of UV light transmitted is reduced because of absorption by the DNA. During electrophoresis the regions of DNA are detected on-line using a UV-sensitive charge coupled device (CCD). As the absorption coefficient is proportional to the mass of DNA the technique is inherently quantitative. The mass of DNA in a region of the gel is approximately proportional to the integrated signal in the corresponding section of the CCD image. This system currently has a detection limit of less than 1.25 ng compared with 2-10 ng for the most popular conventional technique, ethidium bromide (EtBr) staining. In addition the DNA sample remains in its native state. The removal of the carcinogenic dye from the detection procedure greatly reduces associated biological hazards.