مقایسه خاموش کننده های فلورسنت و غیرفلورسنت روش Real-time PCRدر پلات های آمپلیفیکیشن استاندارد ژن دلتا-6 دساتوراز در کشت سلولی رده PANC-1

Background: employing non-fluorescent quenchers in real-time pcr is appropriate for gene expression examination.objective: the objective of this study was to compare real-time pcr fluorescent and non-fluorescent quenchers in standard amplification plots of delta-6 desaturase gene in panc-1 cell line culture. methods: this analytical study was conducted in the reference laboratory affiliated to qazvin university of medical sciences in 2012. human pancreatic cancer cells (panc-1) were cultured in 75 cm2 flasks, 3x106 cells were seeded in 6-well plates and were treated with specific intracellular signaling drugs. changes in expression of delta-6 desaturase gene were examined with fluorescent and non-fluorescent quenchers using real-time pcr in equal conditions, separately. data were analyzed using student t-test and gene expression results were analyzed using ??ct method with the assumption of 100% efficiency.findings: employed quenchers showed different absorption of the fluorescence emitted by the reporter and caused different results in real-time pcr. using non-fluorescent quencher, the amplification plot was more precise and its baseline was lower. therefore the signal to noise ratio (s/n) was decreased. also, the threshold cycle (ct) value was lower because of increased tm (melting temperature) and decreased non-specific bindings.conclusion: with regards to the results, non-fluorescent quencher is more appropriate compared to fluorescent quencher and can be a better alternative for current quenchers especially in allelic discrimination and snp (single nucleotide polymorphism) studies.