[A new method for EMSA by modifying DIG high prime DNA labeling and detection starter Kit II].
Gel retardation, also named electrophoretic mobility shift assay (EMSA), is a useful tool for identifying protein-DNA interactions. Typically, 32P-labeled DNA probes used in EMSA are sensitive. However, it relies on the handling of hazardous radioisotopes, and is not easily quantified. Recently, some successful cases have been reported using non-radio labelled probes instead of radiolabelled probes in EMSA. The method is rapid, convenient, and safe, but it depends on a very expensive kit. In this study, we offered a new method performing EMSA by modifying DIG High Prime DNA Labeling and Detection Starter Kit II (Rohe). Firstly, the prepared labeled probe was introduced the EcoR I stick in the end of probe for 3'-end labeling, and then was performed the probe labeling and detecting the signals of EMSA with the relatively cheap DIG High Prime DNA Labeling and Detection Starter Kit II Rohe. By adjusting the experiment parameters, the successful result was obtained. The present study provides a successful example and method for modifying DIG High Prime DNA Labeling and Detection Starter Kit II.
Qi
Xiao-Ting
XT
Department of Biology, Graduate University of Chinese Academy of Sciences, Beijing 100049, China. qixiaoting@mail.cnu.edu.cn
Chai
Xiao-Qing
XQ
Liu
Jing
J
Chai
Tuan-Yao
TY
D017931:DNA Primers / Q000737:chemistry* / Q000235:genetics; D004076:Digoxigenin / Q000737:chemistry*; D024202:Electrophoretic Mobility Shift Assay / Q000191:economics / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry; D011933:Reagent Kits, Diagnostic / Q000191:economics; D013194:Staining and Labeling / Q000379:methods*; D014026:Nicotiana / Q000737:chemistry / Q000235:genetics
D017931:DNA Primers; D005456:Fluorescent Dyes; D011933:Reagent Kits, Diagnostic; D004076:Digoxigenin