Lots of helpful information. I have a question: why do you have double bands in 1D amplitude which I saw that in my experiments, like, at 4:40 you have double grey droplets bands? what causes that? non-specific binding of your primers , Probe Cross-reactivity or anything else.
I believe this is fluorescence bleed-through from FAM channel. This phenomenon is often the case with HEX channel, when performing such duplex (FAM+HEX) assays
Lots of helpful information. I have a question: why do you have double bands in 1D amplitude which I saw that in my experiments, like, at 4:40 you have double grey droplets bands? what causes that? non-specific binding of your primers , Probe Cross-reactivity or anything else.
I believe this is fluorescence bleed-through from FAM channel. This phenomenon is often the case with HEX channel, when performing such duplex (FAM+HEX) assays
Very good explanation
TM
63°