A new minor groove binding asymmetric cyanine reporter dye for real-time PCR.
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 micro M 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.
Bengtsson
Martin
M
Department of Chemistry and Bioscience, Chalmers University of Technology 41296 Göteborg and TATAA Biocenter, Medicinaregatan 9E, 413 90 Göteborg, Sweden.
Karlsson
H Jonas
HJ
Westman
Gunnar
G
Kubista
Mikael
M
D000818:Animals; D052160:Benzothiazoles; D002232:Carbocyanines / Q000737:chemistry*; D004247:DNA / Q000737:chemistry* / Q000235:genetics; D003959:Diamines; D005456:Fluorescent Dyes / Q000737:chemistry; D006801:Humans; D051379:Mice; D009690:Nucleic Acid Conformation; D009930:Organic Chemicals; D016133:Polymerase Chain Reaction / Q000379:methods*; D011726:Pyridinium Compounds / Q000737:chemistry*; D011804:Quinolines; D013050:Spectrometry, Fluorescence; D013844:Thiazoles / Q000737:chemistry*
C473386:4-((3-methyl-6-(benzothiazol-2-yl)-2,3-dihydro- (benzo-1,3-thiazole)-2-methylidene))-1-methyl-pyridinium iodide; D052160:Benzothiazoles; D002232:Carbocyanines; D003959:Diamines; D005456:Fluorescent Dyes; D009930:Organic Chemicals; D011726:Pyridinium Compounds; D011804:Quinolines; D013844:Thiazoles; C098022:SYBR Green I; D004247:DNA