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制备型琼脂糖凝胶电泳过程中DNA的保护:抵御紫外线诱导的损伤

Protection of DNA during preparative agarose gel electrophoresis against damage induced by ultraviolet light.

作者信息

Gründemann D, Schömig E

机构信息

University of Heidelberg, Heidelberg, Germany.

出版信息

Biotechniques. 1996 Nov;21(5):898-903. doi: 10.2144/96215rr02.

Abstract

Preparative gel electrophoresis of double-stranded DNA usually includes staining the gel with ethidium bromide followed by illumination with ultraviolet (UV-B) light. In this report, DNA isolated from agarose gels was found to be a poor substrate for in vitro transcription, transformation of E. coli and PCR. Inhibition was not caused by enzyme-inhibiting impurities in the agarose gel, but was induced by a standard transilluminator fitted with 312-nm tubes. Interestingly, it was possible to protect the DNA against UV damage by the addition of cytidine or guanosine to the electrophoresis buffer.

摘要

双链DNA的制备性凝胶电泳通常包括用溴化乙锭对凝胶进行染色,然后用紫外(UV-B)光照射。在本报告中,发现从琼脂糖凝胶中分离出的DNA是体外转录、大肠杆菌转化和聚合酶链反应(PCR)的不良底物。抑制作用不是由琼脂糖凝胶中的酶抑制杂质引起的,而是由装有312纳米灯管的标准透射仪诱导的。有趣的是,通过在电泳缓冲液中添加胞苷或鸟苷,可以保护DNA免受紫外线损伤。

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