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使用 FAM 标记的 DNA 寡核苷酸进行 RNA 电泳迁移率变动分析。

Using FAM labeled DNA oligos to do RNA electrophoretic mobility shift assay.

机构信息

Key Laboratory of MOE for Plant Developmental Biology, College of Life Sciences, Wuhan University, 430072 Wuhan, People's Republic of China.

出版信息

Mol Biol Rep. 2010 Jul;37(6):2871-5. doi: 10.1007/s11033-009-9841-7. Epub 2009 Sep 26.

Abstract

The electrophoretic mobility shift assay is a useful tool to identify proteins and nucleic acids interactions. Traditionally, the nucleic acids fragments are end-labeled with (32)P. We present here the use of fluorescent methodologies for studies of RNA in place of radioactivity. The method is highly sensitive and quantitative with the use of an infrared fluorescence imaging system. Fluorescently labeled primers can be used to monitor protein-RNA interactions by fluorescent mobility shift assays. The simplicity and validity of this approach may have more advantages than that of previous methods that traditionally used hazardous radioisotopes. This method was successfully tested in the study of the interactions between MS2 Coat Protein and its RNA target.

摘要

电泳迁移率变动分析是一种用于鉴定蛋白质与核酸相互作用的有用工具。传统上,核酸片段采用(32)P 进行末端标记。我们在此介绍使用荧光方法学来替代放射性物质研究 RNA。该方法使用红外荧光成像系统具有高灵敏度和定量性。通过荧光迁移率变动分析,荧光标记的引物可用于监测蛋白与 RNA 的相互作用。与传统上使用危险放射性同位素的先前方法相比,这种方法具有更多的优势,简单且有效。该方法已在 MS2 外壳蛋白与其 RNA 靶标之间相互作用的研究中成功进行了测试。

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