Quantitative gel electrophoresis of DNA: resolution of overlapping bands of restriction endonuclease digests.
Quantitative gel electrophoresis of DNA: resolution of overlapping bands of restriction endonuclease digests.
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DNA 定量凝胶电泳：限制性内切核酸酶消化的重叠条带的分辨率。
DOI：
10.1016/0003-2697(91)90165-p
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发表时间：
1991
影响因子：
2.9
通讯作者：
Sutherland,JC
中科院分区：
文献类型：
--
作者：
Ribeiro,EA;Sutherland,JC
采用电荷耦合器件检测器的电子成像系统极大地促进了由限制性内切酶完全消化并在电泳凝胶中分离的DNA分布的准确定量。这种定量允许提取亲本分子种类和由限制性内切核酸酶产生的片段的长度的更精确值。我们描述了一种方法，该方法确定了含有两个或多个子片段的条带中的组分数量，所述子片段具有相似的长度，因此不能通过凝胶电泳解析。我们还报告了一个受约束的反卷积程序的基础上，在一个终端消化的限制性内切酶，解决了每个重叠的物种在凝胶上的复合带中的分布，从而增加了分配给每个长度的准确性的子分子的数量相等。
Accurate quantitation of the distribution of DNA digested to completion by a restriction endonuclease and separated in an electrophoretic gel is greatly facilitated by electronic imaging systems employing charge-coupled device detectors. Such quantitation permits the extraction of more accurate values of the lenghts of both the parent molecular species and the fragments produced by a restriction endonuclease. We describe a method that determines the number of components in bands containing two or more daughter fragments that are of similar length and hence not resolved by gel electrophoresis. We also report a constrained deconvolution procedure based on the equal number of daughter molecules in a terminal digestion by a restriction endonuclease that resolves the distribution of each of the overlapping species in a composite band on a gel and hence increases the accuracy of the lengths assigned to each.