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Efficiencies did not change significantly between different cDNA samples (Additional file 11: Table S2).
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Experiments were repeated three to five times with different cDNA samples to confirm the findings.
The mean value obtained for 5 different cDNA samples was +1.63 ΔΔCT for the CCND1 assay.
Different cDNA samples were amplified with primers for 16S rRNA, and the cDNA samples were normalized by differential dilutions according to quantity of 16S rRNA products.
After reverse transcription cDNA was purified using Qiaquick PCR purification kit (Qiagen), according to manufacturer's instructions, and equivalent concentrations of the three different cDNA samples were pooled, due to the lack of sample cDNA.
Two replicates were performed on two different cDNA samples, one-day-old and one-to-seven-days-old adults.
Three replicates were performed on two different cDNA samples (one-to-seven-days-old adults) for both RpL8 and OR7 normalized samples.
For qPCR at least two different cDNA samples were generated and analyzed.
Shown are the mean normalized values of three independent experiments, each in duplicates, using different cDNA samples.
β-actin was used as a housekeeping gene to evaluate and compare quality of different cDNA samples.
To compare data from different cDNA samples, the SPSII CT values were normalized against the expression of TaPP2A.
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