Maximise Electroporation Transfection Efficiency and Reproducibility with the CUY21SC Electroporation System and CU902 Polarity Exchanger

Reproducible electroporation and enhanced transfection efficiency depends on strict adherence to the formula:

V = IR

(Voltage = Ampere (I) x Ohm (R)

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Fluctuations in electrical resistance (between electrodes) impact on delivered current values.

Compromised current delivery negatively impacts reproducibility

And, if electrical resistance is different from one electroporation event to the next, then transfection efficiency will also be different (as the corresponding ampere will be different).

For example:

Experiment 1:

Setting 50V, Electric resistance 500 ohm, Ampere 100mA

Experiment 2:

Setting 50V, Electric resistance 5000 ohm, Ampere 10mA
In this example, the transfection results will be different

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For example:

– If the target sample between the electrodes becomes dry, the electrical current will decease because

(generally) drying makes electric resistance higher

– In in utero electroporation, in some cases, this drying indicates experimental failure

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TO CORRECT for these physical experimental factors, the CUY21SC system (prior to delivering the electroporation program) first tests the target sample for R. This then enables precise control of the current setting (I) to ensure delivery of the required V.

Thus, the CUY21 system, is what we call, a current controlled device.

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Competing devices are what we call voltage controlled devices.  Such devices only require the researcher to sets the voltage setting prior to running the electroporation program.  Such devices simply store charge on a capacitor and then release the energy into the sample without measuring the resistance (impedance).  No account is taken of the relevant physical experimental factors and the negative impact they can have on reproducible results if not corrected for.

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In contrast, many competing devices deliver decaying square waves

With the CUY21 electroporation system we have conclusively determined that for specific and sensitive in utero targets:

– voltage must be precisely controlled and

– current below 10mA must be measured very accurately

For example, for optimal gene delivery into single cell or sensitive targets the required voltage is 1V or 2V (and voltage should be set in 0.1 mA resolutions).  This is why the voltage range of the CUY21SC is from 0.1 to 99.9V in 0.1 resolutions.

For most in utero electroporation protocols, voltage control between 20-40V is most often recommended.  To reflect this recommendation, the voltage control of the CUY21SC system under 30V (indeed under 100V) is completely stable. This is not the case for many competing devices.

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.Example CUY21SC Protocol:

Pulse Length:       50ms

Pulse Interval:    100ms

No. of Pulses:           3

Voltage:                  15V

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Electrodes listing by product code

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To further enhance electroporation transfection efficiency we recommend the

CU902 polarity exchanger

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Nepa Gene Electroporation and Cell Fusion Monitoring System:

For your information, this is the Monitoring System that was used to demonstrate the CUY21SC system outputs above.

With the Monitoring system it is now possible to monitor in real time ALL the controllable parameters required for efficient and repeatable Electroporation (and Cell Fusion). REAL TIME measurement of experiment output parameters enables the researcher to quickly amend and optimise experiment settings and TROUBLESHOOT to solutions to those ‘why is it not working as I expected this time’ questions.

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