7900ht qpcr system thermal cycler

Total RNA was extracted from the cells and tissues using the Trizol reagent (Invitrogen). Approximately 1 μg of RNA was used for the reverse transcription reaction using the PrimeScript™ RT reagent Kit with gDNA Eraser (Vazyme). The cDNA was amplified with the following primers: 5′-CACAGACCTGGCCCGTTTT-3′ (forward) and 5′- AGTCCGGCCTTTCTTTTTGC-3′ (reverse) for ING4; 5′-CTTCCAAGTCTGGAGCGATGT-3′ (forward) and 5′-TACCGTCAAAGGGGTATCCAT-3′ (reverse) for MMP-2; 5′-GTACTCGACCTGTACCAGCG-3′ (forward) and 5′-AGAAGCCCCACTTCTTGTCG-3′ (reverse) for MMP-9; 5′-TCTCGACATCGAGGACCCAT-3′ (forward) and 5′-TGGACCAGTCGAAACCCTTG-3′ (reverse) for TIMP-2; 5′-CTGTCTAATGCCCTGGAGCC-3′ (forward) and 5′-ACGCGAGTCTGTGTTTTTGC-3′ (reverse) for VEGF; 5′-CTGGCGCTCAGCCATACAG′ (forward) and 5′-CGCACTTATACTGGTCAAATCCC′ (reverse) for COX-2; 5′-GGTGCCTTTTCACAGGCTC'-3′ (forward) and 5′-GCTGTTCTCATTGGGTGACTC-3′ (reverse) for Sp1; 5′-GCCGGTGCTGAGTATGTC-3′ (forward) and 5′-CTTCTGGGTGGCAGTGAT-3′ (reverse) for GAPDH.
Real time PCR was carried out in triplicate with SYBR Green PCR Master Mix using a 7900HT qPCR system thermal cycler (Applied Biosystems) as described previously [37 (link)]. GAPDH mRNA was used as an internal control for each sample, and the Ct value for each sample was normalized to GAPDH mRNA.

Total RNA were extracted from collected cells by Trizol (Invitrogen) and reverse‐transcribed (RT) using a cDNA synthesis kit (TaKaRa, China) according to the manufacturer's instructions. The expression of the genes encoding AIM2, ASC, caspase‐1 and IL‐1β was quantified by real‐time PCR using 7900HT qPCR system thermal cycler (Applied Biosystems) and SYBR Green system (TaKaRa) normalized by GAPDH expression following the manufacturer's protocol. Primers were used as following in Table 1.