What is Neon transfection system?
The Neon Transfection System offers an innovative electroporation transfection method that utilizes a proprietary biologically compatible pipette tip chamber to generate a more uniform electric field for a significant increase in transfection efficiency and cell viability.
What is a good transfection efficiency?
With cationic lipid-mediated transfection, generally 70–90% confluency for adherent cells or 5 × 105 to 2 × 106 cells/mL for suspension cells at the time of transfection provides good results.
What is the difference between electroporation and Nucleofection?
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.
Does Confluency affect transfection?
Cell confluency and replication stage are factors intrinsically linked to each other that affect the efficiency of your transfection protocols. This is because cells that are actively dividing take up DNA more readily than stationary phase cells.
How do you use electroporation machine?
How electroporation works
- Step 1 : Prepare cells. Prepare cells by suspending in electroporation buffer.
- Step 2 : Apply electrical pulse. Apply electrical pulse to cells in the presence of specialized buffer and nucleic acids.
- Step 3 : Return cells to growing conditions.
- Step 4 : Assay cells.
What is flow electroporation?
MaxCyte’s flow electroporation technology uses an electrical charge to produce the reversible permeability of cell membranes. This allows for the transfer of molecules, such as nucleic acids and proteins, into the cells thereby creating cell therapies.
How can I increase my transfection efficiency?
Improving the Success of Your Transfection
- Transfect healthy, actively dividing cells at a consistent cell density.
- Transfect using high-quality DNA.
- Optimize the amount of DNA used to transfect cells.
- Optimize the transfection reagent:DNA ratio.
- Optimize cell number per well when transfected.
What is the difference between lipofectamine 2000 and 3000?
Lipofectamine 3000 reagent yields higher transfection efficiencies than Lipofectamine 2000 reagent when tested in a variety of cell lines.
Is Nucleofection the same as transfection?
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.
How much does a Nucleofector cost?
As nucleic acids are delivered straight into the nucleus, nucleofected cells can be ready for analysis after 2–6 hrs of transfection. The bad part about the system is that, it is very expensive ($22,150 (USD) for commercial and about $10,000 (USD) for academic use).
Where is HEK293 from?
HEK293 is a cell line derived from human embryonic kidney cells grown in tissue culture. They are also known, more informally, as HEK cells. This particular line was initiated by the transformation and culturing of normal HEK cells with sheared adenovirus 5 DNA.
Does pen strep affect transfection?
We have found that antibiotics (pep/strep) do not affect the ability of lipofectamine reagents to transfect cells.
Can We transfect human THP-1 macrophages with electroporation-based nucleofector technology?
Therefore, we present here an efficient, non-viral protocol to transfect human THP-1 macrophages using the electroporation-based Nucleofector technology, which represents an optimized electroporation approach requiring reduced amounts of DNA. Nucleofection is well-suited for sensitive cells such as monocytes and macrophages.
Is it possible to transfect human THP-1 monocytes?
This approach allows for the transfection of cell lines such as human THP-1 monocytes and macrophages and has been successfully applied in the past6-10.
How is transfection of THP-1 cells with plasmids performed?
Transfection of THP-1 cells with plasmids or small interfering RNA (siRNA) is achieved by electroporation using the Lonza Nucleofector technology (Basel, Switzerland).
Can RPMI-1640 medium be used to cultivate THP-1 cells after transfection?
However the RPMI-1640 medium, which is the default medium used for cultivation of THP-1 cells, has proven to be ill-suited for cultivation after transfection as a significant loss in cell vitality was observed.