Yasukochi T, Ooyama A, Kumazawa H, Kawanabe J, Ozawa K, Shibanaka Y, Nakajima M
Tsukuba Research Institute, Novartis Pharma K.K., Ohkubo 8, Tsukuba, Ibaraki 300-2611, Japan.
Nucleic Acids Res Suppl. 2001(1):119-20. doi: 10.1093/nass/1.1.119.
We have made the first apparatus for fluorescent detection, monitoring the hybridization process of fluorescently labeled DNA fragments in a polyacrylamide gel. Using this, the analysis on the thermal denaturation/reassociation process of DNA fragments in the gel was employed, for improving the performance of In-Gel Competitive Reassociation (IGCR) technique, one of genome subtraction methods. We showed that Fluorescence Resonance Energy Transfer (FRET) in the gel occurred by positioning two fluorescent dyes at 3' and 5' ends of DNA fragments. The characterization of fluorescence-labelled fragments in gel and the changes of their fluorescence intensity will be reported.
我们制造了第一台用于荧光检测的仪器，用于监测聚丙烯酰胺凝胶中荧光标记的DNA片段的杂交过程。利用该仪器，对凝胶中DNA片段的热变性/复性过程进行了分析，以改进基因组消减方法之一的凝胶内竞争性复性（IGCR）技术的性能。我们发现，通过将两种荧光染料分别置于DNA片段的3'端和5'端，凝胶中会发生荧光共振能量转移（FRET）。本文将报道凝胶中荧光标记片段的特性及其荧光强度的变化。
