Li L, McGown L B
Department of Chemistry, Duke University, Durham, NC 27708, USA.
Fresenius J Anal Chem. 2001 Feb;369(3-4):267-72. doi: 10.1007/s002160000663.
Commercially available, replaceable sieving matrices and their solvent modulated forms were evaluated for use in on-the-fly fluorescence lifetime detection of dye-labeled DNA fragments in capillary electrophoresis. The fragments were labeled with dyes that can be excited by the 488 nm line of an argon ion laser and have lifetimes in the range of 0.8 ns to 3.8 ns. The sieving matrices and buffer systems included poly(vinylpyrrolidone) (PVP), poly(ethyleneoxide) (PEO), hydroxyethylcellulose (HEC), Tris-borate-EDTA (TBE) and Tris-TAPS-EDTA buffers modified with DMSO and formamide. Selection of the optimal sieving matrix is based on the separation efficiency and the enhancement of lifetime resolution of DNA fragments. Best results for both electrophoretic resolution and lifetime detection were obtained using a poly(ethyleneoxide)/TBE gel buffer in the presence of 10% formamide.
对市售的可更换筛分基质及其溶剂调制形式进行了评估，以用于毛细管电泳中染料标记的DNA片段的实时荧光寿命检测。这些片段用可被氩离子激光的488nm谱线激发且寿命在0.8ns至3.8ns范围内的染料进行标记。筛分基质和缓冲系统包括聚乙烯吡咯烷酮（PVP）、聚环氧乙烷（PEO）、羟乙基纤维素（HEC）、Tris-硼酸-EDTA（TBE）以及用二甲基亚砜（DMSO）和甲酰胺改性的Tris-TAPS-EDTA缓冲液。最佳筛分基质的选择基于DNA片段的分离效率和寿命分辨率的提高。在10%甲酰胺存在的情况下，使用聚环氧乙烷/TBE凝胶缓冲液可获得电泳分辨率和寿命检测的最佳结果。
