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 | M Mehdi | en |
| Author | M Darabi | en |
| Author | SH Biagowi | en |
| Author | R Najafipour | en |
| Author | T NaserpoorFarivar | en |
| Author | M Darabi Amin | en |
| Issued Date | 2014-02-13 | en |
| 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. | en |
| DOI | https://doi.org/ | en |
| Keyword | Real-Time PCR | en |
| Keyword | Delta-6 Desaturase | en |
| Keyword | Cell line | en |
| Publisher | Brieflands | en |
| Title | 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 | en |
| Type | Research Article | en |
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