PANC-1 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

M Mehdi; M Darabi; SH Biagowi; R Najafipour; T NaserpoorFarivar; M Darabi Amin

Journal of Inflammatory Diseases

The Official Journal of Qazvin University of Medical Sciences

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PANC-1 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

Author(s):

M Mehdi,

M Darabi,

SH Biagowi,

R Najafipour,

T NaserpoorFarivar,

M Darabi Amin^,*

Journal of Inflammatory Diseases:Vol. 17, issue 5; 17-24

Article type:Research Article

How to Cite:M MehdiM DarabiSH BiagowiR NajafipourT NaserpoorFarivarM Darabi Aminet al.PANC-1 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.J Inflamm Dis.17(5):e155778.

Abstract

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.

Keywords

Real-Time PCR

Delta-6 Desaturase

Cell line

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