The Ct is good,peak is suitable, the band is seen on gel electrophoresis. but efficiency is inadequate. The instrument shows about 80%.dose any one know why?
Are you talking about amplification efficiency and how you calculate it.
Please attach the amplification curves. Ideally attach the melting curves, too. Otherwise it is impossible to help you.
How are you determining efficiency, and why do you feel it is inadequate?
80% isn't great, but it is workable in most cases - you just need to run more cycles. If your product is long or your primers are not optimal 80% may be as good as you are likely to get.
I agree with the suggestions above: more information is needed. What kind of chemistry do you use? Is it a single peak and a single band on the gel? How do you determine efficiency - in silico from a single amplification curve or using a standard curve?
Low PCR efficiency can be for a number of reasons. Assuming that you have a single peak in the melting curve profile and a single band on the gel and use a standard curve to calculate reaction efficiency, my guess would be that either the template you used for the standard curve degraded or that the dilutions were prepared incorrectly. Alternatively, if you work with difficult samples such as blood or soil, carry-over of PCR inhibitors could also affect the reaction efficiency.
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