Focusing and stabilization of bis-intercalating dye-DNA complexes for high-sensitive CE-LIF DNA analysis

CORC > 生态环境研究中心 > 中国科学院生态环境研究中心 > 环境化学与生态毒理学国家重点实验室

	Focusing and stabilization of bis-intercalating dye-DNA complexes for high-sensitive CE-LIF DNA analysis
	Wang, Zhixin; Wang, Chao; Yin, Junfa; Li, Tao; Song, Maoyong; Lu, Meiling; Wang, Hailin
刊名	ELECTROPHORESIS
	2008-11
卷号	29 期号:22 页码:4454-4462
关键词	CE DNA analysis Intercalating dye LIF On-line focusing
英文摘要	Multiple labeling of nucleic adds by intercalative dyes is a promising method for ultra-sensitive nucleic acid assays. The properties of the fast dissociation and instability of dye-DNA complexes may prevent from their wide applications in CE-LIF nucleic add analysis. Here, we describe an optimum CE focusing method by using appropriately paired sample and separation buffers, Tris-glycine buffer and Tris-glycine-acetic acid buffer. The developed method was applied in both uncoated and polyacrylamide coated fused-silica capillary-based. CE-LIF analysis while the sample and separation buffers were conversely used. The complexes of intercalative dye benzoxazolium-4-pyridinium dimer and dsDNA were greatly focused (separation efficiency: 1.8 million theoretical plates per meter) by transient isotachophoresis mechanism in uncoated capillary, and moderately focused by transient isotachophoresis in combination of field amplified sample stacking and further stabilized by the paired buffer in polyacrylamide coated capillary. Based on the developed focusing strategy, an ultrasensitive DNA assay was developed for quantitation of calf thymus dsDNA (from 0.02 to 2.14 pM). By the use of an excitation laser power as low as 1 mW, the detection limits of calf thymus dsDNA (3.5 kb) are 7.9 fM in concentration and 2.4 x 10(-22) mol (150 molecules) in mass. We further demonstrate that the non-gel sieving CE-LIF analysis of DNA fragments can be enhanced by the same strategy. Since the presented strategy can be applied to uncoated and coated capillaries and does not require special device, it is also reasonable to extend to the applications in chip-based CE DNA analysis.
WOS记录号	WOS:000261741100002
内容类型	期刊论文
源URL	[http://ir.rcees.ac.cn/handle/311016/21950]
专题	生态环境研究中心_环境化学与生态毒理学国家重点实验室
推荐引用方式 GB/T 7714	Wang, Zhixin,Wang, Chao,Yin, Junfa,et al. Focusing and stabilization of bis-intercalating dye-DNA complexes for high-sensitive CE-LIF DNA analysis[J]. ELECTROPHORESIS,2008,29(22):4454-4462.
APA	Wang, Zhixin.,Wang, Chao.,Yin, Junfa.,Li, Tao.,Song, Maoyong.,...&Wang, Hailin.(2008).Focusing and stabilization of bis-intercalating dye-DNA complexes for high-sensitive CE-LIF DNA analysis.ELECTROPHORESIS,29(22),4454-4462.
MLA	Wang, Zhixin,et al."Focusing and stabilization of bis-intercalating dye-DNA complexes for high-sensitive CE-LIF DNA analysis".ELECTROPHORESIS 29.22(2008):4454-4462.