With its superior transfection performance, Nucleofection offers various advantages over traditional electroporation methods: High transfection efficiencies of up to 90% for plasmid DNA and 99% for oligonucleotides, like siRNA. Excellent preservation of the physiological status and viability of transfected cells.
Is Nucleofection the same as electroporation?
Nucleofection is an electroporation-based transfection method which enables transfer of nucleic acids such as DNA and RNA into cells by applying a specific voltage and reagents. Nucleofection, also referred to as nucleofector technology, was invented by the biotechnology company Amaxa.
What is transfection and electroporation?
Electroporation is a physical transfection method that uses an electrical pulse to create temporary pores in cell membranes through which substances like nucleic acids can pass into cells.
How does Lipofection differ from electroporation?
Generally speaking, lipofection is used to transfect attached cells, while electroporation is used for suspended cells. Compared to lipofection, electroporation can cause much more cell damage and the efficiency is usually around 30% in B-cell lines.
What are transfected cells?
Broadly defined, transfection is the process of artificially introducing nucleic acids (DNA or RNA) into cells, utilizing means other than viral infection.
What is flow electroporation?
Flow electroporation is a universal, regulatory-compliant transient transfection technology that provides a practical solution to the time, labor, and cost challenges of developing stable cell lines and baculovirus-based expression while overcoming the flexibility and scalability limitations associated with other ...
What is the overall purpose of Lipofection?
Lipofection, also known as “lipid transfection” or “liposome-based transfection,” uses a lipid complex to deliver DNA to cells. Lipids are a broad class of fat-soluble biomolecules, such as fats, oils, and waxes.
How do transfections work?
Transfection is the process of introducing nucleic acids into eukaryotic cells by nonviral methods. Using various chemical or physical methods, this gene transfer technology enables the study of gene function and protein expression in a cellular environment.
What cells can be transfected?
Transfection terminologyTransfection. Transfection commonly refers to the introduction of nucleic acids into eukaryotic cells, or more specifically, into animal cells. Transformation. Transformation is often used to describe non-viral DNA transfer in bacteria, non‑animal eukaryotic cells, and plant cells. Transduction.
What do you need to always know about electroporation based DNA vaccines?
Studies with electroporation assisted DNA vaccinations have shown that it is possible to obtain both a cellular and a humoral immunological response, and in the preclinical setting the technique has shown promising results. However, the effect in large animals and humans may still be improved.
How is electroporation carried out?
Electroporation is based on a simple process. Host cells and selected molecules are suspended in a conductive solution, and an electrical circuit is closed around the mixture. An electrical pulse at an optimized voltage and only lasting a few microseconds to a millisecond is discharged through the cell suspension.
How do you confirm transfection?
Determining the number of positive cells within a transfected cell population can be done through microscopy and flow cytometry. Finally, confirming localization of your protein of interest can be done by microscopy.
What is a good transfection efficiency?
Generally, it is recommended that passaging of cells should take place once or twice a week. The optimal level of confluency before transfection varies depending on cell type and should always be determined ab initio if data cannot be found; very generally, it lies around 70-80%.
How are cells transfected?
Transfection can be carried out using calcium phosphate (i.e. tricalcium phosphate), by electroporation, by cell squeezing or by mixing a cationic lipid with the material to produce liposomes that fuse with the cell membrane and deposit their cargo inside.
What cell type is electroporation used?
The process of introducing foreign DNA into eukaryotic cells is known as transfection. Electroporation is highly effective for transfecting cells in suspension using electroporation cuvettes. Electroporation has proven efficient for use on tissues in vivo, for in utero applications as well as in ovo transfection.