Mm il18 1 sg quantitect primer assay

Total RNA was isolated using the DNA/RNA Shield (Zymo research) and the innuPREP RNA mini kit (Analytik Jena). cDNA was synthesized with innoScipt reverse transcriptase (Analytik Jena). Real time-PCR analysis of IL-15 and IL-18 was performed using innuMIX quantitative PCR (qPCR) MasterMix SyGreen (Analytik Jena). Oligonucleotide sequences were ordered at Biomers as follows: for β-actin, 5′-AAATCGTGCGTGACATCAAA-3′ and 5′-CAAGAAGGAAGGCTGGAAAA-3′; IL-15, 5′-CATTTTGGGCTGTGTCAGTG-3′ and 5′-TCTTCAAAGGCTTCATCTGCAA-3′. For the detection of mouse IL-18 Mm-Il18-1-SG QuantiTect primer assay was purchased from Qiagen. The quantitative mRNA levels were determined by using Rotor-Gene Q series software (Qiagen) and were normalized to the β-actin mRNA expression levels.

Real time-PCR analysis for the quantification of IFN-α, IL-12, IL-15, and IL-18 mRNA was performed using innuMIX quantitative PCR (qPCR) MasterMix SyGreen (Analytik Jena). Oligonucleotide sequences (Biomers) for tested genes were as follows: for β-actin, 5′-AAATCGTGCGTGACATCAAA-3′ and 5′-CAAGAAGGAAGGCTGGAAAA-3′; for IFN-α, 5′-ATGGCTAGGCTCTGTGCTTTCC-3′ and 5′-AGGGCTCTCCAGACTTCTGCTCTG-3′; for IL-12 p40, 5′-ACAGCACCAGCTTCTTCATCAG-3′ and 5′-TCTTCAAAGGCTTCATCTGCAA-3′; and for IL-15, 5′-CATTTTGGGCTGTGTCAGTG-3′ and 5′-TCTTCAAAGGCTTCATCTGCAA-3′. The Mm-Il18-1-SG QuantiTect primer assay (Qiagen) was used. The quantitative mRNA levels were determined by using Rotor-Gene Q series software (Qiagen) and were normalized to β-actin mRNA expression levels.