changing the syber green reagent - (Dec/13/2007 )
I'm doing real time PCR using the syber green method for a long time. Recently I got to try a new syber green reagent from a different company (Kapa biosystems).
The new reagent gave lower Ct's than the old reagent (Thermo scientific- Absolute blue) - I repeated the experiment with four samples.
My questions are- Are there any other parameters that I need to check before deciding on which reagent to use? how to I decide which of the reagents is better?
Reproducibility (are you always getting similar Cts when you repeat your pcr with the same samples in a different run, and are your duplicates Cts close) and sensitivity when using low amount templates are important factors too...