The Universal Cell Transfection Reagent is a polymer-based reagent suitable for transfecting adherent cells, suspension cells, and primary cells. It can effectively deliver nucleic acids such as DNA, mRNA, and siRNA into a variety of cell lines, including suspension cell lines like Expi293, CHO-S, K562, and THP-1, as well as adherent cell lines such as HEK-293T, 4T1, A549, and PC, along with ocular and neural primary cells. This product has advantages including simple operation, high protein expression level, low cytotoxicity, and good cell universality. It is supplied as a sterile liquid.
Product Components and Storage Conditions:
T751608
| Components
| 100μL
| 500μL
| 1.5mL
| Storage
|
T751608A
| Universal Cell Transfection Reagent
| 100μL
| 500μL
| 1.5mL
| -20℃
|
T751608B
| Transfection Buffer
| 2mL
| 10mL
| 30mL
| -20℃
|
Experimental Materials:
• This product contains the Universal Cell Transfection Reagent and the Transfection Buffer .
• High-purity nucleic acid stock solution (0.5–5 μg/μL).
• Serum-free medium (e.g., O778393 Optimal-MEM Reduced Serum Medium (with Phenol Red) ).
Quality Control:
• The shelf life is 1 year when stored at -20°C.
• Strict quality inspection has confirmed that this product is free from biological contamination.
Precautions:
• Prepare the Universal Cell Transfection Reagent-nucleic acid complex at room temperature, with an incubation time of 15 minutes. For transfection, use serum-free medium (e.g., O778393 Optimal-MEM Reduced Serum Medium (with Phenol Red)) and enzyme-free tubes.
• After transfecting with this product for 4-6 hours, add an equal volume of fresh complete medium.
• For first-time use, the ratio of nucleic acid concentration to the Universal Cell Transfection Reagent should be optimized to achieve the highest transfection efficiency. Test using the product dosage within the recommended range.
• For transfection, use healthy cells that are properly preserved and regularly passaged, and ensure the medium is free from bacterial, fungal, or mycoplasma contamination. If the cells are recently revived from liquid nitrogen-frozen stocks, they should be passaged at least twice before transfection.
• This product should be stored hermetically at -20°C. Before use, place it at room temperature to thaw for 10 minutes in advance.
• After using the transfection reagent, immediately replace the cap tightly and store it hermetically at -20°C.
Example of transfection effect:
Figure 1. Cell positive rate and green fluorescent protein expression level 48 hours after delivering pDNA in Exipi293 and CHO-S suspension cells using the Universal Cell Transfection Reagent and competing products, as well as gene silencing efficiency 48 hours after delivering GFP-siRNA in GFP-HeLa cells (24-well plate, 1 μg of pDNA transfected per well).
Figure 2. Gene silencing efficiency 48 hours after delivering GFP-siRNA in GFP-HeLa cells using the Universal Cell Transfection Reagent and competing products. (24-well plate, 1.5 μg of siRNA transfected per well; the dimmer the GFP fluorescence, the lower the GFP expression level and the higher the silencing efficiency.)
Figure 3. Fluorescence expression 48 hours after delivering pDNA in adherent cells such as 293T and 4T1 using the Universal Cell Transfection Reagent and competing products (24-well plate, 1 μg of pDNA transfected per well).
Figure 4. Fluorescence expression 48 hours after delivering mRNA in adherent cells such as 293T and DC using the Universal Cell Transfection Reagent and competing products (24-well plate, 1μg of mRNA transfected per well).
Transfect cells according to the table below. The volume of each reaction mixture refers to the volume for a single well, with pipetting differences taken into account. Calculate the volumes for other wells in proportion.
[T751608] Universal Cell Transfection Reagent.pdf
Universal Cell Transfection Reagent----Detailed Instructions for Transfection Steps (Taking 24-well Plate as an Example)
1. Before use, place the Universal Cell Transfection Reagent at room temperature to thaw 10 minutes in advance.
2. Take 20 μL of Transfection Buffer (pH ~5) into a 1.5 mL centrifuge tube, and add 1 μg of nucleic acid to it to obtain the nucleic acid dilution.
3. Take 1 μL of Universal Cell Transfection Reagent into another 1.5 mL centrifuge tube (The dosage of Universal Cell Transfection Reagent can be optimized within the range of 0.7–1.5 μL to achieve the best transfection effect).
4. Transfer the nucleic acid dilution from Step 2 into the centrifuge tube containing Universal Cell Transfection Reagent from Step 3, mix thoroughly until uniform, and incubate at room temperature for 15 minutes.
5. Mix the incubated Universal Cell Transfection Reagent-nucleic acid complex with 480 μL of serum-free medium (e.g., O778393 Optimal-MEM Reduced Serum Medium (with Phenol Red)), then add the mixture to the cells.
6. After incubating the cells at 37°C for 4–6 hours, add an equal volume of complete medium. Continue incubating the cells at 37°C for 2–4 days, then analyze the transfected cells.
Universal Cell Transfection Reagent----Detailed Instructions for Transfecting Suspension Cells (Taking a 125mL Culture Flask and 30mL Culture System as Examples)
1. Before use, place the Universal Cell Transfection Reagent at room temperature to thaw 10 minutes in advance.
2. Take 1.2 mL of Transfection Buffer (pH ~5) into a 1.5 mL centrifuge tube, and add 60 μg of nucleic acid to it to obtain the nucleic acid dilution.
3. Take 60 μL of Universal Cell Transfection Reagent into another 1.5 mL centrifuge tube (The dosage of Universal Cell Transfection Reagent can be optimized within the range of 32–90 μL to achieve the best transfection effect).
4. Transfer the nucleic acid dilution from Step 2 into the centrifuge tube containing the Universal Cell Transfection Reagent from Step 3, mix thoroughly until uniform, and incubate at room temperature for 15 minutes.
5. Directly add the incubated Universal Cell Transfection Reagent-nucleic acid complex into the cell culture flask, incubate the cells at 37°C for 2–4 days, and then analyze the cell transfection status.
