Manual

Product Number: TR9810

Shipping and Storage

This product is transported at low temperature and stored at -20℃, with a validity period of 1 year to avoid repeated freezing and thawing. If it cannot be used all at once, 0.1ml can be divided into small portions for 0.2cm electric rotary cups, and 0.25ml can be divided into small portions for 0.4cm electric rotary cups.

Description

EZ Transfer electrotransfection reagent (electrotransfection buffer) is used to efficiently and low toxicity transfect nucleic acids (DNA, siRNA, etc.) into difficult to transfect cells. EZ Transfer is compatible with various conventional electrometers, including Lonza Amaxa®Nucleofector®,Bio-Rad®Gene Pulser and Harvard-BTX® electroporators. EZ Transfer can be used for exponential decay and square wave form electroporation experiments.

Important optimization tips

1 Lonza (Amaxa) cell nuclear power transfer instrument

When using the EZ Transfer transfection reagent with the Lonza (Amaxa) cell nuclear transfection instrument, please follow the operating guidelines and optimization plan of the Lonza (Amaxa) cell nuclear transfection instrument, just replace the electroporation mixture with EZ Transfer.

2 Cell density during electroporation

Determine the optimal cell density for each cell type to maximize the efficiency of electroporation. In the final system containing electroporation reagents, the cell density during electroporation is generally within the range of 1-10×10⁶ cells/ml. For suspended cells, it is best to have a high cell density close to 10×10⁶ cells/ml. For adherent cells, the recommended cell density is 1-5×10⁶ cells/ml. You can refer to Tables 1 and 2 to understand the cell density at which EZ Transfer reagents start.

3 Nucleic acid purity and concentration

3.1 Do not use DNA purified solely by ethanol precipitation method. The residual salt in the ethanol precipitation method can cause a change in current and have a negative impact on electroporation.

The recommended concentration range for DNA extraction is 1-5 mg/ml, dissolved in pure water or TE. The use of high concentration DNA may lead to uneven mixing with cells. A lower nucleic acid concentration may dilute the electroporation mixture.

3.2 SiRNA uses high-purity and sterile siRNA.Determine the optimal siRNA concentration for electrotransfection. It is recommended to conduct gradient experiments with different concentrations of siRNA within the final concentration range of 250-750nM.

4 Pulse condition optimization

The EZ Transfer electrotransfection reagent can be used for both exponential decay electroporation experiments and square wave electroporation experiments. Some types of cells are better transfected in the form of square wave pulses, while others respond better to exponential decay forms.

5 Cell incubation time after electrotransfection

There is an optimal incubation time for each cell type after electrotransfection. For plasmid electrotransfection, the optimal incubation time is generally 4-48 hours, and the specific optimal time needs to be adjusted according to the experimental purpose, plasmid properties, and half-life of the expressed protein.

6 The issue of cell death in electrotransfection

When using EZ Transfer, due to the damage of electroporation to cells, according to experience, the transfection effect is best when the mortality rate after transfection is around 50%.

Related products

TR2000: Used for transfecting DNA into HEK293T cells.