Generally, for G-Quadruplex labelled with FAM and TAMRA FRET pairs it is a convention to monitor the changes in FAM emission (530 nm) to construct the FRET melting curves. Is there any specific reason for that? Can TAMRA (acceptor) emission not be monitored?
If you do not want to measure the emmision of TAMRA, I think you could measure the decreasing of emmision of FAM for FRET efficiency.
Dear Guobing Xiang,
I want to measure TAMRA emission only for FRET assay. However in most papers FAM emission is monitored , so I wanted to know if it is okay to use TAMRA emission.
Sneha,
In theory, of course you can measure the emission of TAMRA.
In case of Fam/Tamra, if the two dyes are far apart, you can not see any Tamra emmision, so it is better to measure the emmision change of Fam May be it is better to have reading than reading 0 for some data, I think.
Fam and Tamra have to be close enough for FRET to take place to see emission of TAMRA.
Just mine 2 cent, FYI only!
FRET pair FAM/Quencher(acceptor) is also commonly used, Quencher will not have emision, so you can only measure the emission change of FAM.
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