Expression of BCL2 (an apoptosis gene) in HeLa cells was analyzed by one-step RT-PCR on the LightCycler 2.0. Duplicate reactions were run using 10-fold RNA dilutions (100 ng to 0.1 ng). Unlike [B] the instrument-dedicated kit from Supplier R (which was used according to the fast cycling protocol), the [A] QuantiFast SYBR® Green RT-PCR Kit provided specific amplification (single peak in melting curve analysis) and sensitive quantification (lower CT values that are evenly spaced from each other).
The QuantiFast SYBR® Green RT-PCR Kit provided accurate quantification of HSP89 (a heat shock protein) transcript over a wide dynamic range in a one-step RT-PCR reaction on the Applied Biosystems 7500 Fast System. Reactions were run in triplicate using 10-fold dilutions of in vitro transcript (107 down to 10 copies).
The QuantiFast SYBR® Green RT-PCR Kit reduces total RT-PCR run time by up to 60% in real-time one-step RT-PCR on standard cyclers (40 cycles without melting curve analysis; comparison with standard QIAGEN real-time PCR kits). L: LightCycler 2.0; A1: ABI PRISM 7900; A2: Applied Biosystems 7500; A3: ABI PRISM 7000.