Science with J

You have amplified and measured the Ct of your serial cDNA dilutions... Now what? To validate your primer couple  (i.e., to measure their efficiency) , you have probably been recommended the serial dilution method. What does the serial dilution method for validating primers entail? This method involves making dilutions of your typical control sample (e.g., 1 in 4 , 1 in 16 , 1 in 64 , 1 in 256 , 1 in 1024 ) and mixing it with your typical Master mix and your favorite primer concentration.  This way you can confirm in silico whether the amplification pattern of your primers in successively diluted samples fits a trend line. DNA strand on which the PCR reaction is carried out. How to measure the efficiency of your primer couple using the serial dilution method If you measured Ct values ​​from at least 3 replicates of each dilution, calculate the Ct average of the 3 replicates for each dilution . Consider your 1 in 4  sample as a hypothetical cDNA concentration value of 10...