I am checking GFP fluorescence level in heme sensor. 200 ul of sample was loaded in black flat bottom 96 well plate in H1 Synergy machine. Temperature was 30 degree.
I am using S. cerevisiae WT in W303 background as control.
Are you sure it is not due to the background noise or autofluorescence? Actually, I have faced the same problem with eGFP constructed plasmids, but I was aware that it was originally caused by high background since you know that the 488 channel is highly luminous.