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Nucleic Acids Research, 2003, Vol. 31, No. 8 e45
© 2003 Oxford University Press

A new minor groove binding asymmetric cyanine reporter dye for real-time PCR

Martin Bengtsson, H. Jonas Karlsson, Gunnar Westman and Mikael Kubista

Department of Chemistry and Bioscience, Chalmers University of Technology 41296 Göteborg and TATAA Biocenter, Medicinaregatan 9E, 413 90 Göteborg, Sweden

*To whom correspondence should be addressed. Tel: +4631 773 3926; Fax: +4631 773 3948; Email: mikael.kubista{at}tataa.com

The minor groove binding asymmetric cyanine dye 4-[(3-methyl-6-(benzothiazol-2-yl)-2,3-dihydro- (benzo-1,3-thiazole)-2-methylidene)]-1-methyl-pyridin ium iodide (BEBO) is tested as sequence non- specific label in real-time PCR. The fluorescence intensity of BEBO increases upon binding to double-stranded DNA allowing emission to be measured at the end of the elongation phase in the PCR cycle. BEBO concentrations between 0.1 and 0.4 µM generated sufficient fluorescence signal without inhibiting the PCR. A comparison with the commonly used reporter dye SYBR Green I shows that the two dyes behave similarly in all important aspects.


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