: Reporter Assays
Q: How do I choose between the Cignal DNA-based Reporters and the Cignal Lenti Reporters?
A: The DNA-based Reporters are designed for cells that are amenable to transfection. Qiagen recommends a minimum transfection efficiency of 25 – 30%. If the cells are refractory to transfection then the Cignal Lenti reporters should be selected.
Q: Can I transform the Cignal DNA-based Reporters?
A: No. The Cignal DNA-based Reporters are not designed for bacterial transformations.
Q: Can I make stable cell lines with the Cignal DNA-based Reporters?
A: No, the Cignal DNA-based Reporters are not designed to generate stable pathway sensor cell lines. However, the Cignal Lenti Reporters can be used to generate stable pathway sensor cell lines.
Q: What transfection reagents are compatible with the Cignal DNA-based Reporters?
A: Any transfection reagent that yields 25-30% transfection efficiency in the cell line of interest is suitable for use with Cignal DNA-based Reporters.
Q: Can users insert custom transcriptional regulatory elements or promoters into the Cignal Reporters?
A: No. The Cignal Reporters do not currently support this functionality.
Q: The pathway reporter luciferase activity values are greater than the positive control, is there a problem?
A: No. The positive control only serves as a constitutively-active reporter enabling the user to assess if the transfection/transduction was successful, and should not be used for direct comparison to the pathway reporters.
Q: The pathway reporter luciferase activity values are less than the negative control, is there a problem?
A: No. The negative control only serves as a non-inducible reporter, but should not be used for direct comparison to the pathway reporters.
Q:What is MOI?
A: MOI is an abbreviation for Multiplicity of Infection or the number of viral particles exposed to a cell.
Q:How many experiments can I do with the Cignal Lenti Reporters?
A: This depends on the Multiplicity of Infection (MOI) required for a specific cell line. The lower the MOI the more cells that can be transduced. Therefore, to predict how much Cignal Lenti Reporter is required an MOI must be determined.
Q: What MOI should I use for my cells?
A: This has to be empirically determined by the user. By using a positive control Cignal Lenti Reporter (GFP
) the user can setup parallel transductions with increasing amounts of virus to identify an MOI that yields a robust signal.
Q: What cell culture plates are used for the Cignal Arrays?
A: Each Cignal Array has the DNA reporters "printed" on the bottom of a 96-well cell culture microplate. The microplates have white walls with clear flat bottoms. The wells have a poly-D-lysine surface.