Real-time quantitative PCR analysis of pro-inflammatory gene expression in Human embryonic kidney cells 293 (HEK293)

To assess the role of 8-oxoguanine DNA glycosylase 1 (OGG1) on TNF induced pro-inflammatory gene expression. We used Qiagen Human Inflammatory Cytokines & Receptors PCR Array (PAHS-011Z) to quantitate inflammatory gene expression dependent on OGG1. Triplicate cultures of cells (n = 3; -/+OGG1 SiRNA, -/+ Negative control SiRNA) were TNF exposed for 30 minutes or no TNF exposed, RNAs were isolated, pooled, cDNA were synthesized using Superscript® III First Strand Synthesis System (Invitrogen), mixed with equal amounts of 2X iTaq Universal SYBR Green Supermix (Bio-Rad) and 20 μl of reaction mixture was added to each well of the PAHS-011Z array. The qRT-PCR was performed using an ABI PRISM® 7000 Sequence Detection System as recommended by SA Biosciences.