Volume 54, Number 9, 1507-1514, DOI: 10.1007/s11434-009-0107-5

mRNA detection in living cell using phosphorothioate-modified molecular beacon

HongXing Tang, XiaoHai Yang, KeMin Wang, WeiHong Tan and Wei Li

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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.

Keywords  molecular beacon - phosphorothioate modifying - living cell imaging - mRNA

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)

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