How does a Nucleofector work?
Table of Contents
How does a Nucleofector work?
The Nucleofector Technology uses a specific combination of optimized electrical parameters and cell type-specific solutions which enables transfer of a molecule directly into the cells’ nucleus.
Is Nucleofector 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 are plasmids in transfection?
Plasmids are small circular DNA molecules that naturally occur in bacteria, and are actually used by the bacteria to transfer genetic information. The mechanism of adding a DNA plasmid into a mammalian cell is known as plasmid transfection.
How much DNA do you need for PEI transfection?
In general, use 1 µg of DNA per 1 ml of culture to be transfected. PEI and DNA should each be diluted into 1/20 of the total culture volume before being combined.
What is 4D Nucleofector?
The 4D-Nucleofector X Unit is one of the four functional modules of the 4D-Nucleofector System. It supports Nucleofection of various cell numbers (2 x 104 to 2 x 107) cells in different formats. There are cell type-specific Optimized Protocols or recommendations available in our knowledge database.
How are plasmids transferred into bacteria?
Bacteria can take up foreign DNA in a process called transformation. Transformation is a key step in DNA cloning. It occurs after restriction digest and ligation and transfers newly made plasmids to bacteria. After transformation, bacteria are selected on antibiotic plates.
How do you make PEI for transfection?
Preparation of PEI Stocks
- Dissolve 100 mg in 100 mL sterile ddH2O.
- Stir while slowly adding HCl to pH 7.0.
- Mix for 10 minutes and then recheck pH.
- Filter sterilize through 0.22 um filter.
- Aliquot 500 uL to 1000 uL and store in -80C.
- Thawed solution can be stored at 4C for up to 2 months, label a tube when thawed.
How much plasmid do I need for transfection?
Optimal amount of Universal Transfection Reagent used depends on cell type and is generally 1 – 3 µL per ug of plasmid DNA.
Can plasmid DNA be transferred by transduction?
Transduction involves the transfer of either a chromosomal DNA fragment or a plasmid from one bacterium to another by a bacteriophage. Conjugation is a transfer of DNA from a living donor bacterium to a living recipient bacterium by cell-to-cell contact.
