Advanced  Transfection Reagent

(AD600025/AD600075/AD601000)

I-Application：

-The highest transfection efficiency in kinds of cell types.

-Advanced DNA RNA Transfection Reagent™ can be used for DNA transfection, siRNA transfection and co-transfection in various eukaryotic cell lines.

-Advanced DNA RNA Transfection Reagent™ can be used for DNA transfection, siRNA transfection in various primary cells.

II-Product Description：

Advanced DNA RNA Transfection Reagent™ is a newly developed reagent for transfection of DNA and siRNA into eukaryotic cell lines and various primary cells.

Storage Conditions:  4°C (Don’t Freeze)

Period of Validity: 2 years

III-Transfection requirement

Plasmid DNA:           200ng/ul-2ug/ul;

Dissolved in ddH₂O or ultra-pure water;

Endotoxin removed;

siRNA:                    20 uM/L

IV-Transfection Procedure

1. Planting cells on the cell culture plate one day in advance, the cell confluence degree should be 60-80% at the time of transfection, and the cell state should be kept. Transfection should be carried out in good condition.

2. Complex preparation: Nucleic acid was directly mixed with transfection reagent according to 1:1 relationship, then use a pipette to blow 10-15 times to mix. After incubation at room temperature for 10-15 minutes, the transfection complex was prepared. During preparation of the complex, no liquid residue was ensured on the tube wall.

3. Add transfection complex to the cell culture plate and mix gently, place in the CO₂ incubator and continue to culture. 

Transfection gradient 

 V- Cell viability analysis:

Cell growth was assessed using the Cell Counting Kit (K009, ZETA LIFE Inc.) , which is an indirect measure of cell viability.

VI-The important guidelines：

-Plasmid DNA must be dissolved in ddH₂O. If it was dissolved in Buffer, the transfection efficiency will decrease by 70%, even lead to the failure of transfection.

-Plasmid DNA must be de-endotoxin, otherwise the transfection efficiency will decrease by 70%,  even lead to the failure of transfection.

-No reagent else can be used to dilute nucleic acid or transfection reagent during preparation of transfection complex. Just mix nucleic acid and transfection reagent directly according to 1:1 relationship. Otherwise, the transfection efficiency will be decreased by 80%, even lead to the failure of transfection.

-After checking and mixing with transfection reagent, the pipette was used to blow and suck 10-15 times to mix sufficiently and ensure that there was no liquid residue in the tube wall.

-The nucleic acid was mixed with the transfection reagent and incubated at room temperature for at least 10-15 minutes.

-Add the complex to the cell culture plate and mix gently, place the plate in the CO₂ incubator and continue to culture.

-In the whole process of transfection experiment, cells can be cultured in complete medium instead of using serum-free medium.

-If white or black round particles are observed under microscope after transfection, they will be new material particles of transfection reagent.

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