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Standard curves for PCR efficiency.


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#1 scistudent

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Posted 24 November 2009 - 05:34 AM

Hi Just wondering is 4 points enough for a standard curve to get the efficiency of primers/probes in a qPCR reaction? Im doing each point in triplicate and: no dilution, 1/2, 1/10, 1/100 do I need to do another 1/500 dilution or are the four points enough? (I'm trying to fit as many one to one plate as possible).
Thanks!

#2 ivanbio

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Posted 24 November 2009 - 07:17 AM

Hi scistudent

A 4-point standard curve is be enough to get the efficiency of your reaction. There is nothing special about a 5-point standard curve other than the fact that it covers a broader range of template concentrations and if one dilution does not work you can always remove it from the analysis.

Good luck

Ivan
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#3 scistudent

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Posted 24 November 2009 - 11:40 AM

Ok great thanks! now I can fit all 6 on my plate woohoo,
Thank you!!

Hi scistudent

A 4-point standard curve is be enough to get the efficiency of your reaction. There is nothing special about a 5-point standard curve other than the fact that it covers a broader range of template concentrations and if one dilution does not work you can always remove it from the analysis.

Good luck






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