High efficiency and high viability without special buffers
The most popular transfection procedures all require specialized chemicals, which are costly—as much as $8, $12, or even $16 or more per sample! These additives can have unwanted effects on cells; at the very best, they create another parameter to be dealt with. Though poking holes in cell membranes has been done for many years, no protocols exist for high efficiency transfections without the use of such reagents. That is, not until the NEPA21. Nepa Gene has created THE most sophisticated apparatus for delivering DNA and RNA to cells—and all that’s needed are the electrodes. With a combination of low voltage short-and-long pulses and reversing polarities, a huge variety of cells, tissues, and even whole organisms can be transfected with amazingly high efficiencies. This patented pulsing technology has been shown to be both superior to and far more flexible than other techniques. Relying upon unique non-capacitor driven pulsing, the NEPA21 works at low voltages to deliver perfect square waves with no degradation. Results with the NEPA21 are often far better than those with cheaper models, especially when it matters most. And because everything works at low voltages, cytotoxic effects are nearly eliminated! Read more about the secrets behind NEPA21’s proprietary pulsing technology below.
In vitro transfection of suspension, adherent, and primary cells
The NEPA 21 utilizes one of two electrode types to deliver nucleic acids to live cells in culture. The first type, inexpensive electroporation cuvettes, can be used on suspension cells without the need for expensive buffering conditions with unknown additives. Just add cells and pulse–then save time and money. For adherent cells, use a special cell-culture-plate electrode for 2-dimensional electro-kinetic transfer of RNA and DNA. . In defiance of conventional wisdom, the patented electric pulsing pricks channels not just in the plasma membrane, but the nuclear envelope as well—allowing DNA to be transported into the cell nucleus for efficacious transfection results with primary cells, stem cells, ES cells, and iPSCs alike.
In vivo transfection in mice and rats
The NEPA21 works with a plethora of electrodes that puts Dr. Frankenstein to shame. These specialized tools can be used to gently pulse organs and tissues in live animals, carrying DNA and RNA molecules directly into the cells that make up these tissues. These tools allow previously difficult or impossible transfection experiments to be executed, opening new avenues of scientific inquiry. Transfections have been positively performed on muscle, skin, liver, kidney, testis, ovary, brain, retina, cornea, and others in mice and rats. Click on the following tissue types to see the specific electrodes and protocols designed for each: brain, retina, muscle, skin and testis. And if you need to study tissue outside of the organism, the NEPA21 is also amenable to ex vivo transfer of nucleic acids to a variety of tissue types such as brain, cochlear tissue, or organoids.
In utero, ex utero and in ovo transfections for developmental studies
Nepa Gene has developed tools and techniques for transfecting embryonic tissues–or even whole embryos!–in mice, rats and chickens. Whether it’s specific regions in a mouse embryonic brain (click for a movie), chicken cortex, or chicken gastro regions, we can provide a protocol and an electrode set. The flexibility is almost limitless; these electrodes have been adopted for use with a variety of other animals and plants including honey bee, xenopus, and even plant seeds. Give us a call to discuss the options in more detail.
Click here to see available electrodes for in vivo, in utero, in ovo and ex vivo applications.
This figure compares actual output voltages of samples (in 2 mm electroporation cuvettes) as measured on a Tektronix oscilloscope. The NEPA21 (Panel A) and a premium-brand electroporator (Panel B) performed identical pulsing parameters under varying levels of resistance. All other electroporation systems use capacitor-driven pulsing. This technology can work well under low ionic conditions, as represented by the high resistance (1000 ohm) at the top of panel B. But only the NEPA21’s proprietary design can maintain true “square wave” voltage deliveries under lower resistance (200 and 50 ohm) conditions. This is representative of electroporation in saline conditions (such as PBS), which results in improved cell viability and more consistent transfection efficiencies.
Using 2 mm cuvettes, 5 µg of pCAGGS-eGFP was used to electroporate 1 x 106ES cells in suspension. The results combined much higher cell viability with a 5-fold greater transfection efficiency (68% vs 14%) using the NEPA21 (column A) vs a popular competing instrument (column B).
The NEPA21 Transfecton System’s 4-step pulse with voltage decay results in higher transfection efficiency and lower cell damage without the use of special buffers.
Step 1 – Poring Pulse: High voltage, short duration, multiple pulses, voltage decay.
The poring pulse is for forming small holes in the cell membrane with minimal damage.
Step 2 – Poring Pulse Reverse Polarity: This increases efficiency with adherent cells and other types of transfections.
Step 3 – Transfer Pulse: Low voltage, long duration, multiple pulses, voltage decay.
The transfer pulse is for delivering the target molecules (DNA, RNA, etc.). Like low voltage electrophoresis, these extra-long pulses carry charged molecules into cells with minimal damage.
Step 2 – Transfer Pulse Reverse Polarity: This increases efficiency with suspension cells, adherent cells, and other types of transfections by 10-30%.
Viability – 80%
Transfection Efficiency – 83%
Primary Cell Viability (V) and Transformation Efficiency (TE) with the NEPA21 Electro-Kinetic Transfection System
Data below shows disease-specific iPSC generation (disease: LQT) – transfection of multiple episomal plasmids into B cells
NEPA21 Electro-Kinetic Transfection System
Popular Electrodes - Please see our electrode page for full menu
C115CB-2 – 2mm Female to 4mm Male plug & Cover banana plug cable, red & black, 2 meter$149.00 Add to cart
C118 – 2 socket, 2mm Female to 1 socket, 2mm Male (monopole) plug cable, red & black$89.00 Add to cart
In Vitro Electroporation (suspension cells)
Adherent Cell Electroporation (not trypsinized)
In Vivo Zygote Electroporation
CUY501P1-1.5 – 20mm length platinum plate electrodes on glass slide, 1mm gap, 1.5mm height (5-20ul volume)Request Quote
In Utero Embryo Electroporation
|# pulsing phases||Up to 4|
|Types of pulses||Poring and Transfer|
|# of poring pulses||0-9|
|# of transfer pulses||0-99|
|Total energy readout||Yes, in Joules|
|Power Requirements||AC100-240V 50/60Hz|
|Size||346(W) x 330(D) x113(H) mm|