Gene Expression Quantification by qRT-PCR

The qRT-PCR assay has been used for identification of gene alterations in previous studies [44 (link),45 (link)]. Thus, we used this approach to verify our results about Microarray (item 2.2.2.2). Table 1 shows the primer sequences that were employed. To complete the qRT-PCR experiment, we extracted total RNA from each sample using the Qiagen RNA extraction kit, followed by cDNA production using the iScript cDNA synthesis kit with SYBR green supermix (Bio-Rad, Hercules, CA, USA), as directed by the manufacturer. qRT-PCR was used to quantify mRNA using Rotor-Gene RG-300 from Corbett research [46 (link)]. All investigations were carried out in triplicate by three different biological groups.