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#1
08-30-2011, 06:51 PM
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 High Fluorescence using SYTO9 Dye Hi Everyone, I am having trouble with very high background fluorescence when using intercalating dyes. I am designing a high resolution melt experiment and I have been using the recomended concentration of MeltDoctor (SYTO9 green) at 1 uL/reaction with a reaction volume of 25 uL. The cycle threshold often times, when autoanalyzed will come up at 60,000-1,000,000. Is this normal? I am used to using a taq-man probe assay where the cycle threshold will be around... 0.2! I have not attempted to decrease dye concentration yet because I have not optimized other important components (primers and MgCl first). Any information would be great because I am not having any luck finding info about this on the web. Brittany  Last edited by brittanycronk; 08-30-2011 at 06:52 PM.  |
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| dye , fluorescence , high , syto9 |
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