Volume 17, Issue 5 (Dec-Jan 2013)                   2013, 17(5): 17-24 | Back to browse issues page

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Mehdi M, Darabi M, Biagowi S, Najafipour R, NaserpoorFarivar T, Darabi Amin M. Comparison of Real-time PCR fluorescent and non-fluorescent quenchers in standard amplification plots of delta-6 desaturase gene in PANC-1 cell line culture. Journal of Inflammatory Diseases. 2013; 17 (5) :17-24
URL: http://journal.qums.ac.ir/article-1-1484-en.html
1- , mdarabiamin@hotmail.com
Abstract:   (7015 Views)
Abstract AIM: The present study was undertaken to determine the difference between florescence and non-florescence quenchers and their efficiency in Real-time PCR. MATERIALS AND METHODS: PANC-1 cells were cultured in 75cm flasks, 3x106 cells were seeded in 6-well plates and treated with specific signaling drugs, their effect on the expression of delta6 desaturase gene were examined with Real-time PCR in equal conditions, once with florescence and another time with non-florescence quencher. RESULTS: different quenchers absorbances of the fluorescence emission from the reporter, cause different results in Real-time PCR. The amplification plot is more precisely when using non-florescence quencher and its base line is lower. Therefore the signal to noise ratio(S/N) decrease. Also non-florescence quencher with raising the melting temperature cause to decrease in non-specific matching and this led to lower CT ratio. CONCLUSION: Data showing increased performance of using non-florescence quencher compared to florescence quencher and it can be a better replace for common quencher specially in Allelic discrimination and SNP(single nucleotide polymorphism) studies.
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Type of Study: Research | Subject: Genetics

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