Enter the Ct values for 1:10 and 1:100 dilutions into the calculator to determine the primer efficiency. The efficiency is calculated as a percentage.
Primer Efficiency Formula
The following formula is used to calculate the primer efficiency:
E = 10^{(-1 / ΔCt)} - 1Variables:
- E is the primer efficiency
- ΔCt is the difference in threshold cycle (Ct) values between the 1:10 and 1:100 dilutions
To calculate the primer efficiency, raise 10 to the power of the negative inverse of the difference in Ct values between the 1:10 and 1:100 dilutions, then subtract 1 and multiply by 100 to get the percentage.
What is Primer Efficiency?
Primer efficiency is a measure of how effectively a primer set amplifies a target DNA sequence in a polymerase chain reaction (PCR). It is an important parameter in quantitative PCR (qPCR) as it affects the accuracy of the quantification. High primer efficiency indicates that the primers are well-designed and that the PCR conditions are optimal for the amplification of the target sequence.
How to Calculate Primer Efficiency?
The following steps outline how to calculate the Primer Efficiency.
- First, determine the Ct value for the 1:10 dilution.
- Next, determine the Ct value for the 1:100 dilution.
- Calculate the difference in Ct values (ΔCt) between the two dilutions.
- Use the formula E = 10^(-1 / ΔCt) – 1 to calculate the primer efficiency.
- Convert the efficiency to a percentage by multiplying by 100.
- After inserting the variables and calculating the result, check your answer with the calculator above.
Example Problem:
Use the following variables as an example problem to test your knowledge.
Ct value for 1:10 dilution = 15
Ct value for 1:100 dilution = 20
