Primer integrity and specificity analysis. (A) 2% agarose gel electrophoresis of control 8093 RT/PCR products using gapdh, 18 s rRNA, and cassini shows a single band for each target gene. (B) Real time RT/PCR on RNA isolated from non-transfected MEFs (control), MEFs transfected with an EGFP-cassini construct or with empty EGFP-C1 plasmid. Values (mean ± SD of triplicate real-time PCR) are expressed as percent of gapdh. (C, D) Derivative dissociation curves from real-time RT/PCR on mouse (C) and human (D) RNA confirm single product amplification. The cassini amplicon shows a single peak for both murine and human RNA with a Tm of ~76°C. Gapdh primers peak at Tm ~85.5°C. ***p < 0.001 EGFP-cassini compared to EGFP-C1 and control.
Arutyunyan et al. BMC Genomics 2012 13:418 doi:10.1186/1471-2164-13-418