DNA transfection Reagent-Low toxicity, High efficiency
Transfection is the process of inserting genetic material, such as DNA and double stranded RNA, into mammalian cells. Specifically, the insertion of DNA into a cell enables the expression, or production, of proteins using the cells own machinery. On the other side, insertion of RNA into a cell is used to down-regulate the production of a specific protein by stopping translation. While the site of action for transfected RNA is the cytoplasm, DNA must be transported to the nucleus for effective transfection. There, the DNA can be transiently expressed for a short period of time, or become incorporated into the genomic DNA, where the change is passed on from cell to cell as it divides.
Bioquochem has optimized the formula of DNA transfection Reagent- TRANS LT (Lower toxicity, Higher efficiency) to increase the efficiency of the transfection and to minimize cell toxicity.
- -Seed cells until desired cell density in culture plates (6/12/24/48/96-well) or 60/ 100 mm plates
-Let the reagents reach room temperature before use.
- 1. First, add in a tube X µl of medium (medium + Reagent A from the next step) must be accordingly with column number 4 of the Table 1.
2. Then, add in the same tube X µl of Reagent A. Stir mixture. Not vortex.
3. Incubate for 5-15 minutes at RT or 37°C. Longer incubations may adversely affect transfections. This solution is called diluted transfection reagent.
4. After that, dilute DNA into Reagent Plus and then add X µg of DNA to the diluted transfection reagent.
5. Incubate for 10-60 minutes at 37ºC.
6. After that, add transfection mixture (DNA+diluted transfection Reagent) into cell medium and shake
7. Finally, visualize/ analyze the transfected cells after 1-7 days at 37ºC.