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PCR efficiency on 2 machines comparable?


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#1 hianghao

hianghao

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Posted 17 June 2011 - 02:13 AM

If i switch the qpcr cycler to another cycler, do i have to redo the standard curve to get the amplification efficiency? Assuming that my reaction conditions are all the same. I am aware that data from 2 machines cannot be compared directly. What i expect is that since all condition r the same, the efficiency may be affected only by machine (since pipetting error cant b avoided), and degree of changes on the effect will be the same for all primer sets. And since i am doing relative quantification, the changes will be canceled out during the calculation. IS this correct?
I am asking since there is a new qpcr cycler in my lab and the old one that i m using will be transferred back to its institute next week (it was borrowed). But i still have 1 sample to run but wil only manage to get it by the end of this year. And i dont want to spend 8 (targets) X 15 rxns to do the standard curve for my primer due to budget constrain.

In the calculation of relative quantification (Pfaffl 2001), the E value is replaced by E in what form? Lets say E=100%, in the formula we put E as 100 or 1 or 2?

Edited by hianghao, 17 June 2011 - 02:13 AM.





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