What is Transfection?

Transfection is the process by which foreign nucleic acids (DNA or RNA) introduced into eukaryotic cells by non-viral methods.

Using various physical or chemical techniques, DNA or RNA transfer technology enables the study of gene function and protein expression in a cellular environment.

Principle of Transfection

TurboFect in vitro Transfection Reagent is a sterile solution of a proprietary cationic polymer in water.

The cationic polymer forms stable, positively charged complexes with negatively charged DNA.

These complexes protect DNA from degradation and facilitate gene delivery into eukaryotic cells.

TurboFect is ideal for transfection of a variety of cells, such as adhernt cells, suspension cells, including primary and difficult-to-transfect cells.

It can be performed in the presence or absence of serum.

TurboFect demonstrates superior transfection efficiency and minimal toxicity when compared to lipid-based or other polymer-based transfection reagents.

Created with BioRender

Transfection Requirements

Sterile:

1. TurboFect in vitro Transfection Reagent

2. 6 well plates

3. Microtips

4. Growth medium

5. Cells (A549 Cell line)

6. Trypsin EDTA

7. DNA

Non sterile:

8. 70% IPA

9. Cotton balls

Transfection Procedure

1. Prior to the day of transfection, 5 x 104 adherent cells were plated in each well in a 12 well plate containing 1 ml of growth medium.

2. 1μg of DNA was diluted in 100μl of serum free DMEM or RPMI media

3. 2μl of turbofect reagent was added to the diluted DNA (2μl/1μg of DNA) and mixed immediately by pipetting or vortexing and it was kept for incubation for 15-20 minutes at room temperature

4. 100μl of this turbofect/DNA mixture was added dropwise to each well without removing growth medium from the cells

5. Plate was gently rocked to achieve even distribution of the complexes.

6. It was further incubated at 37 ̊C in CO2 incubator

7. After 24 hours, treatment with selection antibiotic (Geneticin) was done at concentration of 400μg/ml.