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Inhibition in qPCR

Inhibition in qPCR - Real-Time PCR and Quantitative PCR Forum

Inhibition in qPCR - Real-Time PCR and Quantitative PCR Forum


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  #1  
Old 03-19-2011, 06:33 AM
Pipette Filler
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Default Inhibition in qPCR



Hi all,

Im having trouble with my qPCR assays. It seems like I'm getting a inhibitor in some of my samples when I'm producing cDNA. I use Invitrogen kits to DNase treat my samples then produce cDNA and sometimes after this the samples are fine, but other times the standard curves I make from these sample are completely off. The first few dilutions won't amplify properly and only by the 4th or 5th dilution do I get a normal sigmoidal curve. Its not due to inhibition in the original sample because I have produced cDNA from the same RNA sample and once its been fine, but the second time it wasn't. I can't work out what is causing this because I'm only adding reagents from the invitrogen kits. Does anyone have any idea of what could be going wrong?
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Old 07-03-2011, 10:13 PM
Pipette Filler
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Default Re: Inhibition in qPCR

Why you treat your samples by DNAse????

Please try to explain more your problem! Because i have a prob with qRT-PCR inhibitor! But when i diluted them, it will be right!
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