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Weird qPCR curves


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4 replies to this topic

#1 cells

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Posted 26 July 2011 - 05:13 PM

Hi, can anybody look at my qPCR curves attached. I'm wondering why it keeps going up instead of reaching a plateau..the melt curve looks fine and I haven't run the product on a gel but is it simply the case that I haven't run enough cycles yet for the plateau to be reached?

Thanks.

Attached Thumbnails

  • pcr curves.jpg


#2 Adrian K

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Posted 26 July 2011 - 07:01 PM

Hi,
I am not expert in this field but...
did you add too many primers? what are the primers molarity you put in and what is the concentration?
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#3 Trof

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Posted 27 July 2011 - 06:33 AM

You don't need to run to plateau. The important part of the curve is the exponential phase. Sometimes the curves don't reach plateau, the reason probably is that the reaction is slowed down by insufficiency of some precursor which isn't completely depleted so the reaction doesn't stop, just continue on very slowly.
But that's really not important.

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#4 cells

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Posted 29 July 2011 - 08:14 PM

Thanks for the responses...yea, I'm not going to worry about it too much:)

#5 mhong

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Posted 02 August 2011 - 04:14 PM

These look normal to me. Perhaps you're more used to looking at the the y-axis in log scale.




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