What is an electroporation cuvette?
Ideal for microbiological studies, the electroporation cuvettes transform bacteria, yeast, mammalian, and plant cell samples by applying an electric field and introducing genes. Featuring the necessary aluminum electrode plates on the sides, the tissue culture filled containers prevent static.
How do you reuse electroporation cuvettes?
Reusing Electroporation Cuvettes
- Rinse the cuvette five times with purified water.
- Fill the cuvette with 0.2M HCl and allow to stand for 10 minutes (but no longer as this will promote corrosion).
- Rinse the cuvette five times with 70% ethanol, again ensuring the chambers are fully washed.
What is needed for electroporation?
Electroporation is based on a simple process. Host cells and selected molecules are suspended in a conductive solution, and an electrical circuit is closed around the mixture. An electrical pulse at an optimized voltage and only lasting a few microseconds to a millisecond is discharged through the cell suspension.
How can you prevent arcing during electroporation?
Store Cuvettes in the Freezer: Some researchers swear by having cuvettes as cold as possible. This means storing cuvettes in the freezer until they’re ready for use, keeping them on ice and even icing for a few minutes before electroporation.
Are electroporation cuvettes reusable?
The cuvettes should be washed for reuse. If washed properly, they can be reused up to 10 times. For each cuvette, do the following while waiting for the cells to recover: Squirt 95% ethanol to fill cuvette more than halfway.
What causes arcing during electroporation?
Arcing may occur due to high concentration of salts or air bubbles. It is essential to add recovery medium to the cells immediately after electroporation. One minute delay can cause a 3-fold reduction in efficiency. Cold and dry selection plates lead to lower transformation efficiency.
Why is ethanol 70 ethanol used to wash the cuvettes?
The ethanol rinses away the HCl and sterilises the cuvette but because it is volatile it makes it easy to dry the cuvette in air.
What is an electroporation cuvettes?
Electroporation Cuvettes High-quality, sterile electroporation cuvettes provide consistent pulse delivery to your valuable samples, ensuring reproducible results. Cuvettes are available in three different gap widths (0.4, 0.2, and 0.1 cm) for optimal field strength delivery to a wide range of cell types.
What is a cuvette used for in microbiology?
These cuvettes are for use during the introduction of DNA into bacterial, yeast, and mammalian cells by the process of electroporation. Their features include: This cuvette is recommended for use with all of our electroporation competent cells.
How do you prepare pUC19 for electroporation?
Place electroporation cuvettes (1 mm) and microcentrifuge tubes on ice. As a positive control for transformation, dilute the control pUC19 by 1:5 to a final concentration of 10 pg/μl using sterile water. Heat-denatured ligation reactions can be used for electroporation directly; however, column purification is recommended.
What are the two most important electrical parameters for electroporation?
The two most important electrical parameters for consideration in electroporation are pulse length and field strength. Field strength is defined as volts/centimeter, where V is equal to the initial peak voltage and cm is equal to the measurement of the gap between the electrodes of the cuvette in centimeters.