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-   -   Weird curve with Taqman (http://www.molecularstation.com/forum/real-time-pcr-quantitative-pcr-forum/76346-weird-curve-taqman.html)

PCRgirl 03-18-2011 12:52 PM

Weird curve with Taqman
 
Hi,

I'm doing a real-time to a virus that I tested before with the classical PCR. I'm using taqman probe. In the first attempts there was just a lot of noise. I tried to put more probe and reduce the annealing. Now it seems I have something, but is not a nice curve. The postive samples have a different patern compared to the negative control and other virus strains. I run a gel with the product to check if the primers were ok and I have a strong band with the right size. With this, can I assume that the problem is the probe? (concentration?). And the curve, that is more noise but with a "peak" only to the positive samples is a signal that the system is working and I just need to optimize? I never see a curve like this before.

Thank you

namishu95 03-23-2011 02:58 PM

Re: Weird curve with Taqman
 
since you got a strong band with the right size, it proved that your primers are good, did you quench at 3',
please check your probe sequence, or re-design, or add quencher at internally modified "T".


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