Duol-glo luciferase assay signal - (Oct/14/2005 )
hi,
1st Q:
my reporter plasmid (PGL3 basic + putative promoter) produce low signal, a few time lower than PGL3 basic only. could it be my fragment hindering the basal expression of the firefly on this vector ?? Is that possible that the fragment cloned into the reporter vector lower its expression instead of promoting it???
2nd Q:
Is there any threshold for the transfection efficiency in order for the reading to consider as credible because the tfx effeciency vary a lot frm cell line to cell line. (Renilla signal: Nih-3t3 = 10 million!!, suspension cells =2000-20,0000, losely adherent cells=20,000). Although the renilla signal are tfx control for data normallization, the huge variation of it among cell line make me doubt.
thanks. happy weekend!!
cyk