Transfection Controls, Mediums and Enhancers

Controls

As should be included in every experiment, positive and negative controls provide confidence that downstream results are trustworthy, along with disproving any false positives. There are three controls a researcher should include on every transfection plate: a positive control, negative control and non-treated control.

Positive Controls

A positive control tells the researcher that the system being utilized is working correctly and the delivery conditions are optimal.  The positive control results aid the researcher in understanding intra- and inter-experimental differences.  The ideal positive control contains these characteristics:

    • • Ubiquitous: Ensures the gene is expressed in all cell types and at a constant level
    • • Abundant expression: Enables accurate and easy quantitation of protein or mRNA levels
    • • Non-lethal: Altering expression of the positive control should not alter the phenotype or viability of the cells
    • • Typical positive controls: Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), Cyclophilin B (PPIB), and Lamin (LMNA)

Negative Controls

Negative controls are designed such that they have no known matching target in the cells and are commonly called a scrambled control.  Ideally, negative controls mimic the test article length, G/C content and contain similar modifications.  Negative controls are used for differentiating sequence-specific from non-specific effects.  The negative control samples are handled and analyzed similarly to test article treated samples. The ideal negative control exhibits these characteristics:

    • • Alien sequence: It is designed as an alien sequence with no known target as to not affect mRNA or protein expression levels
    • • Non-lethal: Should not affect cell viability or global gene expression due to non-target design
    • • Typical negative control: scrambled, non-targeting human and mouse nucleic acid typically referred to as an alien sequence

Non-Treated Control

Non-treated controls should include both a mock transfection (i.e. transfection reagent only) and non-treated control. Data generated from the mock and scrambled control helps in understanding experimental toxicity from either the transfection reagent or an alien cargo when compared to the non-treated sample.

These products and services aid researchers in developing optimized transfection conditions:

Other Transfection Controls