Documents & Support

Neon NxT E10, E100, R, and T buffers have the same compositions as Neon E, E2, R, and T buffers, respectively. However, Neon NxT tips and tubes have different designs compared to the Neon tips and tubes, to improve usability, and therefore are not compatible with the Neon Transfection System.

Find additional tips, troubleshooting help, and resources within our Transfection Support Center.

Was this answer helpful?

The siRNA concentration in Neon transfection refers to the siRNA concentration in the culture medium and not to the siRNA concentration in the electroporation content in the Neon tip. For example, if electroporation is performed with the 100 µL Neon tip and the transfected cells are plated in a 24-well plate that contains 500 µL culture medium, the siRNA concentration is measured as the concentration in the 500 µL culture medium and not the concentration in the 100 µL electroporation content.

Find additional tips, troubleshooting help, and resources within our Transfection Support Center.

Was this answer helpful?

For routine cleaning, we recommend cleaning the surface of the Neon device and Neon Pipette Station with 70% ethanol. Do not use harsh detergents or organic solvents to clean the unit. For deeper cleaning, use a broad-range medical disinfectant. Avoid spilling any liquid inside of the Neon Pipette Station. For accidental spiils (e.g., buffer, water, coffee) inside the Neon Pipette Station, disconnect the station from the main device and wipe the station using dry laboratory paper. Invert and leave the station for 24 hours at room temperature for complete drying. Do not use an oven or autoclave to dry the Neon Pipette Station.

Find additional tips, troubleshooting help, and resources within our Transfection Support Center.

Was this answer helpful?

Clean the surface of the Neon NxT Electroporation Device and Neon NxT Pipette Station with 70% ethanol/isopropyl alcohol and wipe. Do not use harsh detergents or organic solvents to clean the units. Avoid spilling any liquid inside of the Neon NxT Pipette Station. If you accidentally spill any liquid (e.g., buffer or water) inside the Neon NxT Pipette Station, disconnect the station from the main device and wipe the station using dry laboratory paper. Invert and leave the station for 24 hours at room temperature for complete drying. Do not use an oven to dry the Neon NxT Pipette Station.

Find additional tips, troubleshooting help, and resources within our Transfection Support Center.

Was this answer helpful?

As few as 10,000 cells may be used with the 10 µL tips, but this may vary depending on size differences among cell types. One of our customers reported successful transfection of 5,000 primary hair follicle cells. As a general rule, try to avoid using low cell densities as this could reduce viability during electroporation. If it is difficult to avoid low cell densities (ex. primary cells), adjust the voltage to optimize for improved viability. Optimal electroporation conditions are cell type-dependent. Avoid antibiotics in the medium and use medium appropriate for your cell type. For helpful cell-specific electroporation conditions, please visit our Neon cell-specific protocols database https://www.thermofisher.com/us/en/home/life-science/cell-culture/transfection/transfection---selection-misc/neon-transfection-system/neon-protocols-cell-line-data.html?SID=fr-neon-3

Find additional tips, troubleshooting help, and resources within our Transfection Support Center.

Was this answer helpful?

The Neon Transfection Tubes are disposable and we recommend using each tube for a maximum of 10 times for the same plasmid/siRNA/cell type, to minimize the possibility of cross-contamination. In addition, we strongly recommend that a new Neon tube be used for a different plasmid DNA/siRNA or cell type, to avoid cross-contamination. If you need extra Neon tubes to accommodate your experiment, they can be purchased separately (Cat. No. MPT100).

Find additional tips, troubleshooting help, and resources within our Transfection Support Center.

Was this answer helpful?

The Neon NxT Electroporation System relies on the same unique and trusted electroporation technology as the Neon Transfection System (Cat. No. MPK5000), but it has new features that make it easier to use. The Neon NxT pulse generator has an improved feedback loop with user interface notification, and ClipTip technology has been incorporated into Neon NxT pipette tips for secure attachment and easy ejection, along with other ergonomic design improvements.

Find additional tips, troubleshooting help, and resources within our Transfection Support Center.

Was this answer helpful?

The shelf life of the Neon kits is 1 year from the date of shipment.

Find additional tips, troubleshooting help, and resources within our Transfection Support Center.

Was this answer helpful?

We recommend reviewing our Neon Transfection System Protocols and Cell Line Database (https://www.thermofisher.com/us/en/home/life-science/cell-culture/transfection/transfection---selection-misc/neon-transfection-system/neon-protocols-cell-line-data.html) to identify a cell type that is similar in tissue origin, to your own cell and try the recommended parameters in the protocol. This is a good starting point, but some optimization may be needed (ex. adjusting voltage). For example, if you have 293 T cells and you find a protocol for HEK293 cells in the Neon Cell Database, you can use the electroporation parameters of HEK293 cells for 293 T cells, since both are derived from human embryonic kidney.
- You can use the pre-programmed 24-well optimization protocol in the Neon device to optimize conditions for your cell type.
- Contact Technical Support at techsupport@thermofisher.com for further discussion.

Find additional tips, troubleshooting help, and resources within our Transfection Support Center.

Was this answer helpful?

Yes. The Neon NxT Electroporation System can be used for electroporation of any RNAi substrate (siRNA, shRNA, miRNA). You can use the same conditions described in the cell type-specific protocol for DNA, or use the pre-programmed 24-well optimization protocols included in the Neon NxT Device.

Find additional tips, troubleshooting help, and resources within our Transfection Support Center.

Was this answer helpful?

The Neon Transfection System has the following unique advantages:

1. Pipette tip chamber design for easy cell handling, uniform electric field, and higher viability due to minimal pH change during electroporation.
2. Single transfection kit (Neon Kit) compatible with all mammalian cell types, including primary and stem cells, thereby avoiding the need to determine an optimal buffer for each cell type. Only two cell resuspension buffers cover all cell types: T buffer for primary T and B cells, PBMCs, monocytes, and bone marrow–derived cells, and R buffer for established adherent and suspension cells as well as primary adherent cells.
3. Easily scalable for small or large cell culture formats. Use as few as 1 X 10E4 or as many as 5 X 10E6 cells per electroporation in a sample volume of either 10 µL or 100 µL.
4. Over 130 validated online protocols, optimized for ease of use and simplicity. Visit our Neon Cell Database (https://www.thermofisher.com/us/en/home/life-science/cell-culture/transfection/transfection---selection-misc/neon-transfection-system/neon-protocols-cell-line-data.html)
5. Pre-programmed with a 24-well optimization protocol to quickly identify best electroporation parameters by cell type.
6. Flexible payload and applications. Delivers DNA, mRNA, siRNA, and proteins.

Find additional tips, troubleshooting help, and resources within our Transfection Support Center.

Was this answer helpful?

Yes, the Neon tip fits into the Digital Bio/NanoEntech pipette.

Find additional tips, troubleshooting help, and resources within our Transfection Support Center.

Was this answer helpful?

Here are possible causes for low transfection efficiency using the Neon device:

1. Sub-optimal electrical parameters
2.Poor plasmid quality such as endotoxin contamination
3 .Plasmid preparation containing high salt
4. Plasmid quantity too high
5. Cells are stressed or damaged
6. Using same Neon tip more than two times
7. Microbubbles in tip, causing arcing

Find additional tips, troubleshooting help, and resources within our Transfection Support Center.

Was this answer helpful?

The E2 buffer has higher osmolarity than E buffer. Higher osmolarity prevents the leakage of electroporation content from the 100 mL Neon tip, which has a aperture hole at the tip end than the 10 mL Neon tip (pore diameter of the Neon tips: 100 mL tip = 2.10 mm; 10 mL tip = 0.65 mm).

Find additional tips, troubleshooting help, and resources within our Transfection Support Center.

Was this answer helpful?

The Neon Transfection system is the second generation of the Microporator (MP-100) from Digital Bio/NanoEntek. Both instruments feature a unique pipette tip electrode chamber instead of a standard electroporation cuvette. The performance of these instruments is comparable, but the Neon Transfection System has the following improved features:

- Improved user touchscreen interface with larger display area
- Most updated firmware (available at Instrument Management: https://www.thermofisher.com/us/en/home/products-and-services/services/instrument-qualification-services/instruments-and-services-portal.html)
- Pre-programmed with one 24-well optimization protocol
- Improved sensor connector design
- >130 validated cell-specific online protocols

We do not carry an adapter to allow compatibility between former and current devices. The kits and components in both systems are exactly identical except that they are branded as Neon in the Neon Transfection System. The old manual can be used for the Neon Transfection System instrument. More information regarding the history of the Neon Transfection System can be found here (https://www.thermofisher.com/us/en/home/life-science/cell-culture/transfection/transfection---selection-misc/neon-transfection-system/history-of-the-neon-transfection-system.html).

Find additional tips, troubleshooting help, and resources within our Transfection Support Center.

Was this answer helpful?