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一种无需预处理或干燥步骤的琼脂糖凝胶中核酸银染方法。

A procedure of silver staining for nucleic acids in agarose gels without pretreatment or drying steps.

作者信息

Prieto C C, Leonardelli R I, Zalazar F E

机构信息

Sistema de Prácticas Finales, Facultad de Bioquímica y Ciencias Biológicas, Universidad Nacional del Litoral, Hospital Dr. J. M. Cullen, Santa Fe, Argentina.

出版信息

Anal Biochem. 1997 Oct 1;252(1):15-8. doi: 10.1006/abio.1997.2288.

Abstract

In this paper, we report a fast, simple, and reproducible staining protocol for nucleic acids in agarose gels with a sensitivity in the order of 10 pg/mm2. It took only three steps: fixation, incubation with silver ions, and development of the gels (total time 50 min). The resulting calibration curves (area vs ng of loaded DNA) after a densitometric scanning of agarose gels stained with this procedure were linear up to 50 ng of double-stranded DNA. We found this method suitable for routine laboratory use and especially appropriate for densitometric analysis due to homogeneous background development. Furthermore, it avoids the pretreatment and/or drying steps proposed by other authors for agarose gels.

摘要

在本文中,我们报告了一种用于琼脂糖凝胶中核酸的快速、简单且可重复的染色方案,其灵敏度约为10 pg/mm²。该方案仅需三步:固定、用银离子孵育以及凝胶显影(总时间50分钟)。用此方法染色的琼脂糖凝胶经光密度扫描后得到的校准曲线(面积与上样DNA的纳克数)在双链DNA达50纳克时呈线性。我们发现该方法适用于常规实验室使用,尤其适用于光密度分析,因为背景显影均匀。此外,它避免了其他作者提出的琼脂糖凝胶预处理和/或干燥步骤。

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