核酸染料GeneGreen可影响DNA琼脂糖凝胶电泳条带的质量

Nucleic acid dye GeneGreen can influence the quality of DNA bands in agarose gel electrophoresis

目的探讨DNA琼脂糖凝胶电泳条带扭曲、"拖尾"的原因。方法采用"胶染法"和"后染法"2种方法,检测含2种核酸染料(GeneGreen和溴化乙锭)的琼脂糖凝胶中3种DNA Marker条带形态。结果采用"胶染法"的方式电泳,与相同浓度的溴化乙锭相比,含1∶10 000和1∶5 000 2种浓度的GeneGreen琼脂糖凝胶中3种DNA Marker条带呈现明显的扭曲和"拖尾"现象。而采用"后染法"的方式后,两种核酸染料所染的琼脂糖凝胶中3种Marker的条带均呈单一、无扭曲和"拖尾"现象。结论核酸染料GeneGreen可以影响琼脂糖凝胶电泳时DNA条带的质量。在排除DNA本身质量、琼脂糖凝胶质量、电泳液质量以及电压等因素后,若采用"胶染法"进行DNA核酸电泳时出现条带扭曲或"拖尾"时,应考虑到核酸染料质量的问题。采用"后染法"或更换核酸染料的种类可改善DNA电泳条带的质量。

Objective To explore the causes of twisted or tailed DNA bands in agarose gels after electrophoresis.Methods Prestained and poststained methods were employed to exam 3 kinds of DNA marker bands in agarose gel with GeneGreen and Ethidium Bromide,respectively.Results Three kinds of DNA marker bands in agarose gel with 1∶10 000 and 1∶5 000 concentration of GeneGreen showed obvious distortions and trailing phenomena compared with the same concentration of Ethidium Bromide when the prestained method was used for electrophoresis,which were improved when the poststained method was used in agarose gel with GeneGreen and Ethidium Bromide,respectively.Conclusion Nucleic acid dye GeneGreen can affect the quality of DNA bands in agarose gel electrophoresis.When the quality of DNA itself,agarose gel quality,electrophoretic fluid quality and voltage are excluded,the quality of nucleic acid dye should be taken into consideration if the bands twisting or tailing occurs when the DNA nucleic acid electrophoresis is carried out by prestained method.The quality of DNA electrophoresis bands can be improved by using poststained method or replacing nucleic acid dye.