盐酸黄连素作为脱氧核糖核酸荧光染料染色效果的研究  

Staining Effect of Fluorochrome Berberine Hydrochloride on Nucleic Acid in Gels

作　　者：杨全玉[1] 李永生[2] 高秀峰[1] 王艳君[1] 金科[1] 赵瑞[2] 

机构地区：[1]四川大学华西基础医学与法医学院,四川成都610041 [2]四川大学化学工程学院,四川成都610065

出　　处：《分析测试学报》2008年第9期968-972,共5页Journal of Instrumental Analysis

基　　金：四川大学振兴计划科研启动基金资助项目(0082204127092)

摘　　要：考察了盐酸黄连素(berberine hydrochloride)的荧光强度与双链DNA的相关性;优选盐酸黄连素在琼脂糖凝胶电泳中的最佳条件;将其染色效果与溴化乙锭(EB)进行比较;并从相对迁移率角度研究盐酸黄连素对DNA凝胶电泳的影响。结果显示,双链DNA对盐酸黄连素有明显的荧光增强作用,荧光强度与dsDNA的浓度正相关;凝胶中10 mg/L的盐酸黄连素可分辨出10 ng的100 bp DNA条带,在完全相同的操作条件下,对于小片段dsDNA,其灵敏度略低于EB。因盐酸黄连素在碱性溶液中带正电荷,使DNA相对迁移率减小,但较染料EB的影响小。该研究为盐酸黄连素在日常分子生物学研究中的应用提供了理论依据。盐酸黄连素可以作为琼脂糖凝胶电泳中DNA的荧光染料,其荧光强度能够满足常规应用,可以替代强致癌性染料EB。The correlation of the fluorescence intensity of berberine hydrochloride （BB） with doublestranded DNA（dsDNA） was examined and the operation conditions for BB in agarose gels electrophoresis were optimized. The staining effect of BB was imaged and compared with that of ethidium bromide（EB）. The results showed that the fluorescence intensity of BB was enhanced significantly by dsDNA and the increased intensity was proportional to the concentration of dsDNA. A nontoxic and simple staining method with fluorochrome BB for the detection of DNA in agarose gels was thus developed. The method can detect as little as 100 bp per band of DNA（10 ng） stained by 10 mg/L BB in gels. For small fragment of dsDNA, the sensitivity of BB was a bit lower than that of EB under the same experimental conditions, since BB was positively charged in alkaline solution leading to the decrease of DNA relative mobility.

关 键 词：盐酸黄连素 核酸荧光染料 琼脂糖凝胶电泳 溴化乙锭 相对迁移率 

分 类 号：O657.32[理学—分析化学] Q523[理学—化学]