Validation of Microarray data by quantative real-time PCR (qRT-PCR)

The expression levels of 12 representative genes were examined by qRT-PCR to validate the microarry data. The primers for qRT-PCR were designed using Primer 5 (S2 Table). The cDNA synthesis was conducted using PrimeScript RT reagent Kit with gDNA Eraser (TaKaRa, Japan). qRT-PCR was conducted on BioRad CFX96 Real-Time System using SYBR Premix Ex Taq (TaKaRa). For each gene/sample combination, three replicate reactions were carried out. In addition, the 16 S rDNA gene was chosen as a reference gene. The qRT-PCR results were processed by “Bio-Rad CFX Manager 3.1” and gene expression ratios from the qRT-PCR were log₂ transformed.