Product Description
Messenger ribonucleic acid (mRNA) initiates protein translation in the cytoplasm without entering the nucleus, enabling faster protein expression than DNA transfection and avoiding the risk of gene integration.
EZ Trans mRNA transfection reagent is a transfection reagent that combines the advantages of cationic polymers and liposomes, synergistically promoting the entry of transfection complexes into cells and their subsequent release in the cytoplasm. It can achieve efficient and low toxicity transfection effects in various cell types.
Order Information
product name | Item number | specifications |
EZ Trans mRNA transfection reagent | AC04L061 | 300 uL |
EZ Trans mRNA transfection reagent | AC04L062 | 1 mL |
Transportation and storage
Blue ice transportation. Stored at 4 ℃, with a shelf life of 6 months- Stored at 30 ℃ to -10 ℃, with a shelf life of 12 months. Note: Avoid repeated freezing and thawing.
Usage (Taking a 24 well plate as an example, please refer to Table 1 for transfection in other culture dishes)
1. Inoculate cells
For adherent cells: On the day before transfection, plate the cells in a ratio of 1-2x105 cells per well, with a density of 60% -80% at the time of transfection.
For suspended cells: On the day of transfection, before preparing the transfection reagent nucleic acid complex, wash the cells 1-2 times with PBS, resuspend the cells in 250 μ L Opti MEM I serum-free medium, and plate the cells in a 24 well plate with a cell density of 60-80%.
2. Prepare transfection reagent mRNA complex
(1) For each well of cells, add 0.5 μ g of mRNA to be transfected into a 1.5mL centrifuge tube, mix 2 μ L of transfection reagent with mRNA, and incubate at room temperature for 3 minutes.
(2) Add 100 μ L Opti MEM I serum reducing medium (serum-free) to the above mixture, gently mix well, and let it stand at room temperature for 30 minutes to form a transfection reagent nucleic acid complex.
(3) After 30 minutes, add 250 μ L Opti MEM I serum reducing medium (serum-free) to the above complex and mix gently.
3. Transfect cells
(1) Wash the cells 1-2 times with PBS and add 350 µ L of transfection reagent mRNA complex to each well of cells.
(2) Cultivate in a 5% CO2 incubator at 37 ℃ for 12 hours without the need to replace the culture medium, and then the transfection effect can be tested. If a longer transfection time is required, 500 μ L of complete culture medium (including serum) can be supplemented 12 hours after transfection, and testing can be conducted 12-36 hours later.
matters needing attention
1. This product is only for scientific experimental research and should not be used in clinical diagnosis, treatment, or other fields.
2. mRNA quality: Please make sure to use high-quality mRNA that is high-purity, sterile, pollution-free, and free of endotoxins. MRNA can be capped (compared to Cap0 structure, mRNA with Cap1 structure is more suitable for mammalian systems), modified with N1 methyl pseudouridine, and added with poly (A) tail to enhance mRNA stability and lifespan.
3. The entire experimental process uses materials that are free of RNAse and pyrogen, such as centrifuge tubes, gun tips, and buffer solutions.
4. Cell conditions: Cell state greatly affects transfection efficiency. The cells to be transfected should be in a good growth state. It is recommended to use cells that are in the exponential growth phase and have a survival rate greater than 90% for transfection. If the cells are recently revived liquid nitrogen frozen cells, please passage them at least twice before transfection.
5. Cell culture medium requirements: The EZ Trans mRNA transfection reagent transfection experiment requires the use of a serum-free transfection system (Opti MEM serum reducing medium is recommended), as serum can affect the formation of complexes and mRNA transfection efficiency.
6. As certain components in certain special culture media may inhibit cationic polymer mediated transfection, it is recommended to test the compatibility of these media with this product to ensure optimal transfection efficiency.
7. Optimization of reagent dosage: RNA concentration and transfection reagent dosage are affected by cell type and other experimental conditions. For initial use, mRNA concentration and transfection reagent dosage should be optimized to achieve maximum transfection efficiency. Normally, the recommended mRNA dosage for a 24 well plate is 0.2-1 μ g, and the recommended transfection reagent dosage is 2-4 μ L. It is recommended to conduct pre experiments with different dosages within the recommended range for the first use to optimize the transfection effect.
Note: The usage amount of this table is for reference only, and the specific usage amount needs to be optimized according to the cell type and other experimental conditions. a: The amount of culture medium required for diluting the transfection reagent mRNA complex; b: Add the amount of transfection reagent mRNA complex to the culture medium.
Related product recommendations
CCK-8 Kit (Cell Counting Kit-8) (Product Code: AC11L054)
EZ Trans Cell Transfection Reagent (High Efficiency) (Product Code: AC04L092)
This product is only for scientific experimental research and should not be used in clinical diagnosis, treatment, or other fields.
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