ROX and Rotorgene - using a mix with reference dye in Rotorgene (Sep/15/2009 )
I was wondering if anybody has ever used a mix containing an internal reference dye with a Corbett Rotorgene or any other machine that does not require the reference for normalization.
Would it affect the Sybr Green fluorescence readings in any way?
Have a good day
hi there.. I have tried ROX mix with Roche Light Cycler 480
The efficiency will decrease and the LOQ obviously will be compromised... In short for a 5 pg sample the Ct (Cp) value will be less than one in which ROX is used (with the same amount)!!!
These systems as I understand are internally calibrated for normalization so dont require ROX, so when we add ROX, insteead of normalizing it adds to the backgound noice!!!
thank you for your reply,
I think it's better not adding things when they are not required..