Bahrami Ahmad R, Dickman Mark J, Matin Maryam M, Ashby John R, Brown Paul E, Conroy Matthew J, Fowler Gregory J S, Rose James P, Sheikh Qaiser I, Yeung Anthony T, Hornby David P
Department of Molecular Biology, The Transgenomic Research Laboratory, Krebs Institute, Firth Court, Western Bank, Sheffield S10 2TN, UK.
Anal Biochem. 2002 Oct 15;309(2):248-52. doi: 10.1016/s0003-2697(02)00306-8.
SYBR Green 1 is an asymmetrical cyanine DNA-binding dye that provides an opportunity for increasing the sensitivity of nucleic acid detection when used in conjunction with gel electrophoresis. In this paper, we summarize the general properties and specific uses of SYBR green 1 in ion-pair reversed-phase denaturing high-performance liquid chromatography (IP DHPLC). We describe several applications for the WAVE DHPLC platform that illustrate the generic potential of such intercalating dyes in mutation detection and gene expression profiling. We show that SYBR Green 1 obviates the need to use end-labeled oligodeoxynucleotides for the sensitive detection of nucleic acids during chromatography. Moreover the incorporation of SYBR Green 1 into samples and elution buffers does not impair resolution and has no significant effect on the retention times of DNA fragments compared with dye-free DHPLC.
SYBR Green 1是一种不对称花青DNA结合染料，与凝胶电泳结合使用时，它为提高核酸检测的灵敏度提供了机会。在本文中，我们总结了SYBR Green 1在离子对反相变性高效液相色谱（IP DHPLC）中的一般性质和具体用途。我们描述了WAVE DHPLC平台的几种应用，这些应用说明了此类嵌入染料在突变检测和基因表达谱分析中的一般潜力。我们表明，SYBR Green 1在色谱分析过程中无需使用末端标记的寡脱氧核苷酸即可灵敏检测核酸。此外，与无染料的DHPLC相比，将SYBR Green 1掺入样品和洗脱缓冲液中不会损害分辨率，并且对DNA片段的保留时间没有显著影响。
