2 replies to this topic

[mengxibio]

what's the function of ROX reference dye?

And what's wrong with the melting curve?

pls watch the pics,

I choose a well,the No.5 sample,show u the melting curve.Why do the rox dye have a peak? Is it the primer dimer?

and the 3.jpg is the total reasult.

thx!

Attached Thumbnails

• 
• 
• 

Edited by mengxibio, 09 August 2011 - 06:40 PM.

[ivanbio]

ROX is used to normalize the signal of your qPCR reactions across the plate. The idea is that if there is more reaction volume in one well compared to the other, or the amount of light captured in one well is higher than another, assuming the reactions across all wells are the same, the amount of ROX will be used by the instrument to normalize this kind of variation across the plate.

From what I can see in your images it seems like you have contamination in your well.

[mengxibio]

ivanbio, on 09 August 2011 - 06:44 PM, said:

ROX is used to normalize the signal of your qPCR reactions across the plate. The idea is that if there is more reaction volume in one well compared to the other, or the amount of light captured in one well is higher than another, assuming the reactions across all wells are the same, the amount of ROX will be used by the instrument to normalize this kind of variation across the plate.

From what I can see in your images it seems like you have contamination in your well.

thx