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TaqMan probe

TaqMan probe - Real-Time PCR and Quantitative PCR Forum

TaqMan probe - Real-Time PCR and Quantitative PCR Forum


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Old 07-15-2011, 02:53 PM
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Default TaqMan probe



I am designing a new assay with a TaqMan probe (5’-FAM TTA TGC AAA “T”GA ARA CCA ATC CCA TTR CAA Spacer-3’, probe is internally quenched with BHQ at “T” residue). It is probably because of AT rich, the performance of assay was bad, the Signal to Noise Ratio was less than 2, any suggestion about this issue, please see below sequence, the residues with Capital letter are actual probe sequence, I extended four bases on 5’ and two bases on 3’
ttc t TTA TGC AAA “T"GA ARA CCA ATC CCA TTR CAA tg
any suggestion?

Last edited by namishu95; 07-15-2011 at 02:54 PM.
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Old 07-20-2011, 02:56 PM
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Default Re: TaqMan probe

guys,

have you seen probe drop down to baseline with high concentractioon DNA/RNA, see attachment, does any one have such experience, how it happened and how to fix? , increase Tm?, GC contain?
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