LightSwitch™ 3´UTR Control Vectors

Synonyms

LightSwitch, Light Switch, reporter assay, reporter gene, luciferase reporter, Renilla luciferase, RenSP, 3′UTR, 3′ UTR, 3UTR, 3 UTR, 3′UTR control, random 3′UTR 4, negative control, siRNA, miRNA, 32020

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Overview

Because no single control can be ideal for all experimental conditions, Active Motif offers a panel of positive and negative control constructs for the LightSwitch™ 3´UTR Reporter Vector. These controls are used to measure background, to optimize experimental conditions and to normalize for non-specific effects associated with a treatment or change in condition. Note that the unique RenSP luciferase technology utilized in the LightSwitch Assay System eliminates the need for co-transfection of a normalizing control in most cases. So, instead of one control construct being co-transfected with every experimental plasmid, different control vectors can be used in your reporter assay experiment, each in its own well, in addition to the constructs being tested. Using the most suitable positive and negative control vectors for your specific experiment will give you the highest degree of confidence in your results.

Information on all LightSwitch 3´UTR Control Vectors is shown under the 3´UTR Controls tab below.

IMPORTANT:  Because all LightSwitch reporter constructs contain the optimized RenSP luciferase gene, you MUST use our LightSwitch Luciferase Assay Reagents to obtain optimal results. (Other luciferase assay reagents are not compatible with RenSP.)

3´UTR Controls

Empty 3´UTR vector: The empty 3´UTR vector contains the RenSP luciferase reporter gene and its constitutive promoter. This construct may serve as a positive control for the transfection because of its promoter. It may also be utilized for creating additional constructs. For a vector map, annotations and primer & restriction sites, please see the Empty LightSwitch™ Reporter Vectors page.

Housekeeping gene 3´UTR vectors: Housekeeping control constructs contain the 3´UTRs for common housekeeping genes cloned downstream of the RenSP luciferase reporter gene.

Random control 3´UTR vectors: Random control constructs contain non-conserved, non-genic, and non-repetitive human genomic fragments.

Figure 1: LightSwitch assay performed using the positive and negative LightSwitch 3´UTR Controls.
Fifty ng of each 3´UTR Control Vector was transfected into HT1080 cells. Because the constitutive promoter driving the RenSP luciferase gene is the same for all vectors, it is expected that all of the vectors will give comparable signals, as this experiment does not include miRNAs to interact with the various 3´UTR regions and change the amount of luciferase produced.
The Y-axis values are shown in log scale. Absolute and relative activity will vary depending on cell line, experimental conditions and transfection reagents.

3´UTRs of housekeeping genes

Catalog No.	Name	Description	Coordinates	Sequence
32013	LightSwitch™ ACTB 3´UTR Control	Beta-actin	chr7–: 5340002 - 5340681	Get Info
32014	LightSwitch™ GAPDH 3´UTR Control	Glyceraldehyde-3-phosphate dehydrogenase	chr12+: 6517534 - 6517833	Get Info
32015	LightSwitch™ LDHA 3´UTR Control	Lactose dehydrogenase A	chr11+: 18385382 - 18386064	Get Info
32016	LightSwitch™ PPIA 3´UTR Control	Peptidylprolyl isomerase A	chr7+: 44614226 - 44616012	Get Info

Random genomic fragments in 3´UTR vector

Catalog No.	Name	Coordinates	Sequence
32017	LightSwitch™ Random 3´UTR Control 1	chr13+: 82190847 - 82193385	Get Info
32018	LightSwitch™ Random 3´UTR Control 2	chr12+: 82120892 - 82121673	Get Info
32019	LightSwitch™ Random 3´UTR Control 3	chr13+: 26144830 - 26145608	Get Info
32020	LightSwitch™ Random 3´UTR Control 4	chr22+: 1645930 - 1646850	Get Info

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