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!
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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
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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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!!
Thank you!!
ivanbio, on Nov 24 2009, 07:17 AM, said:
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
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
