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mRNA detection in living cell using phosphorothioate-modified molecular beacon
Authors:HongXing Tang  XiaoHai Yang  KeMin Wang  WeiHong Tan  Wei Li
Institution:(1) State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Biomedical Engineering Center, and Key Laboratory for Bio-Nanotechnology and Molecular Engineering of Hunan Province, Hunan University, Changsha, 410082, China
Abstract:In this study, GFP mRNA in COS-7 cell and GFP-transfected COS-7 cell was detected in real time using phosphorothioate-modified molecular beacon based on living cell imaging method. Results showed that phosphorothioate-modified molecular beacon still kept the advantages of molecular beacon, such as, excellent selectivity, high sensitivity, and no separation detection. In addition, this modification could significantly increase the nuclease resistance of molecular beacon. Phosphorothioate-modified molecular beacon can efficiently reduce the false positive signal and improve the accuracy of living cell mRNA detection. Supported by the National Key Basic Research Program (Grant No. 2002CB513110), International Technologies Collaboration Program of China (Grant No. 2003DF000039), and National Natural Science Foundation of China (Grant Nos. 90606003 and 20620120107)
Keywords:molecular beacon  phosphorothioate modifying  living cell imaging  mRNA
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