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DNA浓度的变化显著影响随机扩增多态性DNA指纹图谱的可重复性。

Variations in DNA concentrations significantly affect the reproducibility of RAPD fingerprint patterns.

作者信息

Davin-Regli A, Abed Y, Charrel R N, Bollet C, de Micco P

机构信息

Laboratoire d'Hygiène et de Microbiologie, Hôpital Salvator, Marseille, France.

出版信息

Res Microbiol. 1995 Sep;146(7):561-8. doi: 10.1016/0923-2508(96)80562-6.

Abstract

The influence of the DNA concentration was tested using two different primers and nine DNA samples. Major modifications in the DNA banding pattern were apparent between successive dilutions. Such differences could be explained by concomitant changes in three different molecular conditions: the presence of perfect priming sites, the amplification of rare sites and the existence of mismatch annealing events. At low DNA concentrations (less than 1 pg/microliter), molecular events occurred at random and had a direct consequence on the reproducibility of RAPD profiles. At the appropriate DNA concentration (between 100 ng/microliters and 10 pg/microliters), reproducibility was adequate at a given concentration, but RAPD profiles differed from one dilution to another. These observations demonstrate the usefulness of the bis-benzimide method for quantification of DNA extracts.

摘要

使用两种不同的引物和九个DNA样本测试了DNA浓度的影响。在连续稀释之间,DNA条带模式的主要变化很明显。这些差异可以通过三种不同分子条件的伴随变化来解释:完美引物位点的存在、稀有位点的扩增以及错配退火事件的存在。在低DNA浓度(小于1 pg/微升)下,分子事件随机发生,并直接影响RAPD图谱的可重复性。在适当的DNA浓度(100 ng/微升至10 pg/微升之间)下,在给定浓度下可重复性足够,但RAPD图谱在不同稀释度之间有所不同。这些观察结果证明了双苯甲酰亚胺法在定量DNA提取物方面的有用性。

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