the ratio of pGL3 promoter( control) vector in Dual luciferase assay - Dula luciferase assay (Jul/24/2007 )
Could you please tell me the ratio of pGL3 promoter( control) vector used in Dual luciferase assay. I tried different ratio. but I found a quenching problem. and could you please tell me about Dual luciferase kit, which one is better, Roche? or Promge?
-tulip1-
QUOTE (tulip1 @ Jul 24 2007, 05:23 AM)
Could you please tell me the ratio of pGL3 promoter( control) vector used in Dual luciferase assay. I tried different ratio. but I found a quenching problem. and could you please tell me about Dual luciferase kit, which one is better, Roche? or Promge?
I've used pGL4 constructs with pRL (renilla) for normalizing transfection. I started from 2:1 and ended up to 10:1 with success. As far I understand, in order to use dual luciferase kit (I use promega), you need to use two luciferases (pGL=firefly, pRL=renilla) with different wavelenght. You can compare your construct with PGL3 promoter, but both cotransfected with pRL to properly do a DLA assay.
-rulo-