Quantitative fluorescence of DNA-intercalated ethidium bromide on agarose gels.
Quantitative fluorescence of DNA-intercalated ethidium bromide on agarose gels.
复制标题
琼脂糖凝胶上插入 DNA 的溴化乙锭的定量荧光。
DOI：
10.1016/0003-2697(89)90229-7
复制
发表时间：
1989
影响因子：
2.9
通讯作者：
D. Miller
中科院分区：
文献类型：
--
作者：
E. Ribeiro;L. Larcom;D. Miller
分析琼脂糖凝胶上DNA分布的技术通过二维和一维方法进行了检验。结果表明，除非(i)以仔细、可重复的方式进行电泳，（ii）用内部标准校准薄膜，（iii）使用高分辨率密度测定法分析薄膜，以及（iv）使用适当的背景控制来确定整合的基线，否则在此类分析中DNA浓度会出现非常大的误差。二维扫描产生的结果比一维扫描更精确，但在波段相对均匀的情况下，一维分析给出的结果较好。描述了一种确定准确分布的技术。
Techniques for analyzing DNA distributions on agarose gels are examined by both two-dimensional and one-dimensional methods. It is demonstrated that very large errors in DNA concentration occur in such analyses unless (i) the electrophoresis is performed in a careful, reproducible manner, (ii) the films are calibrated with an internal standard, (iii) high resolution densitometry is used for analyzing the films, and (iv) appropriate background controls are used to determine the baselines for integration. Two-dimensional scanning produces more accurate results than one-dimensional scanning, but in cases where the bands are relatively uniform, the one-dimensional analysis gives good results. A technique for determining accurate distributions is described.