Lai et al., 2017_DataSets_Biology letter
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Cp values and priming efficiencies were calculated using the second derivative maximum method (Roche Lightcycler 480; Rasmussen, 2001) and the LinRegPCR software (Ruijter et al., 2009), respectively. Subsequently, relative mRNA expression levels were calculated using the following formula:
Expression of target gene = (Ega^Cpga/Etar^Cptar)
Where ga is the geometric average of the two reference genes; tar is the target gene of interest, E is priming efficiency and Cp is the crossing point.
Since duplicate cDNA syntheses were performed, and each of these were analysed in duplicates in the qPCR analyses, four data points were present for each original sample for each primer pair used, and their means were used in the mRNA expression calculations.
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2017-08-09
