Nucleofector II From Amaxa Biosystems

Transfection is an essential tool for numerous applications, including studies of gene expression. However, transfection of cells with common methods such as calcium phosphate, DEAE-dextran, liposome-mediated transfer, electroporation or viral vectors is limited because of low transfection efficiency and the extended time required for analysis of gene expression. Amaxa Biosystems has a new electroporation-based transfection method that enables the DNA to directly enter the nucleus while cells are stabilized in an electric field by treatment with a specific solution. The method is called “Nucleofection” and the electoporator system is called Nucleofector II. The system has built-in optimized programs for most common cell types. We analyzed the usefulness of Nucleofector II for transfection of a variety of hard-to-transfect cells like CHO, Epithelial cells and SH-SY5Y neuroblastoma cells and achieved high transfection efficiencies (50 -70 % as measured by flow cytometric analysis of GFP +ve cells) with minimal cell death.

The transfection procedure is simple and straightforward with these basic steps: (1) Preparing cells to be transfected with the nucleofection solution appropriate for the cell type, (2) Adding the nucleic acid to the prepared cells and transferring the mixture to a cuvette (provided in the kit), (3) Placing the cuvette in the system, electroporating cells with the optimized program for the cell type, (4) Harvesting cells from the cuvette and dispersing on to media contained in a dish. The whole process can be done in less than 30 minutes.

The good part about the system is that the method is easy, simple (some optimization required with regard to the appropriate amount of DNA and the type of program used for each cell type), sensitive, requires minimal expertise and no prior plating of cells to be transfected is needed. This in contrast to the alternative methods which are limited by the need to plate cells at a particular density 24-48 hours prior to transfection. In addition, these methods are labor and time intensive. Amaxa has nucleofection kits with optimized protocols available for most common cell types. 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). In addition, you have to purchase kits (around $400 for each kit) for each cell type. The expense can further add up if you are trying to transfect a particular cell type that has not yet been optimized by Amaxa because you will have to buy different kits to optimize transfection efficiency yourself.

Overall, my experience with Nucleofector II from Amaxa Biosystem has been very good. We are using this system regularly for our studies to transfect human, hamster and rat cells. Amaxa Biosystems offers an on site demo and gives you the opportunity to keep the demo unit for a couple of weeks to try out. Amaxa’s website also provides you with detail information and a list of easy to follow optimized protocols for a variety of cell types. The bottom line is that the Nucleofector II is the right choice for fast, easy and efficient transfection of hard to transfect cells.