complicated transfection requires professional help
Posted 24 May 2009 - 02:12 AM
I intend to use the dual-luciferase reporter assay for quantitative measurement of gene expression.
I want to produce stably transfected cells that will express both renilla and firefly luciferases under the control of independent promoters and that the cells will also be expressing GFP constitutively (necessary for sorting the cells by FACS).
I badly need help to address two problems:
1) I need an internal control to relate the luciferase expression to that allows stable expression assay (i.e, contains selectable marker and is constitutively expressed)
2) I guess that the excitation of GFP by renilla luciferase might be a problem as the light emitted by renilla reaction is at 480 nm well within the peak for excitation of GFP (470-490 nm). will that actually happen?
Any advice will be highly appreciated
Thanks in advance
Posted 24 May 2009 - 04:29 PM
Posted 27 May 2009 - 08:10 PM