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First of all, I apologize if my English is not so correct!
I have just begun working on absolute Real Time qPCR and I have a problem with DATA analysis; in particular, from the assay, I obtained a standards curve with a good efficiency (slope= -3,3; R2=0.999) but a very low sensitivity (y-intercept = 17). I know that to consider an assay with a good sensitivity, y-intercept has to be between 33 and 37. If it is higher than 37 , it indicates that the amount of template has not been correctly determined (for example because of the deterioration of the template by too many freeze-thaw cycles of the standard), but what about if the value is too less than 33 as in my case (17)? However, this is not a case, I run the same assay for other samples, obtaining similar results of sensitivity and efficiency. What does it mean?
I will appreciate any suggestions, thanks!