Standard curves for PCR efficiency. - (Nov/24/2009 )
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!
-scistudent-
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
-ivanbio-
Ok great thanks! now I can fit all 6 on my plate woohoo,
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
-scistudent-