BOXTO as a real-time thermal cycling reporter dye |
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Authors: | Ashraf I Ahmad |
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Institution: | Department of Chemical and Biological Engineering-Molecular Biotechnology, Chalmers University of Technology, 405 30 Goteborg, Sweden. ashraf.ahmad@molbiotech.chalmers.se |
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Abstract: | The unsymmetrical cyanine dyes BOXTO (4-6-(benzoxazole-2-yl-(3-methyl-)-2,3-dihydro-(benzo-1,3-thiazole)-2- methylidene)]-1-methyl-quinolinium chloride)and its positive divalent derivative BOXTO-PRO (4-(3-methyl-6-(benzoxazole-2-yl)-2,3-dihydro-(benzo-1,3-thiazole)-2-methylidene)]-1-(3-trimethylammonium-propyl)-quinolinium dibromide) were studied as real-time PCR reporting fluorescent dyes and compared to SYBR GREEN I (SG)(2-N- (3-dimethylaminopropyl)-N-propylamino] -4-2,3-dihydro-3-methyl-(benzo-1,3-thiazol-2-yl)-methylidene]-1-phenyl- quinolinium).Unmodified BOXTO showed no inhibitory effects on real-time PCR,while BOXTO-PRO showed complete inhibition. Sufficient fluorescent signal was acquired when 0.5-1.0 meu M BOXTO was used with RotorGene and iCycler platforms.Statistical analysis showed that there is no significant difference between the efficiency and dynamic range of BOXTO and SG.BOXTO stock solution (1.5 mM) was stable at -20 degree C for more than one year and 40 meu M BOXTO solution was more stable than 5xSG when both were stored at 4 degree C for 45 days. |
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Keywords: | BOXTO BOXTO-PRO real-time PCR unsymmetrical cyanine dyes |
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