Description
Transfection reagent for DNA and RNA
GenieFect 2000 Transfection Reagent is a cationic-lipid transfection reagent for efficient delivery of plasmid DNA and RNA, including siRNA, into a broad range of adherent and suspension mammalian cell lines. It provides high transfection efficiency with low cytotoxicity and a simple, serum-compatible protocol.
The reagent readily forms liposome/nucleic acid complexes and releases the nucleic acid rapidly once inside cells, giving excellent transfection performance with very low cytotoxicity. Serum and antibiotics do not affect transfection, so complexes can be added straight to complete medium, with no medium change needed before or after transfection. It also supports co-transfection of siRNA and plasmid DNA. It is suitable for most eukaryotic cells, including 293T, A549, HeLa, SW480, 4T1, Jurkat, NIH3T3, MDA-MB-453, Vero, K562 and Raw264.7. For research use only.
Key Features
| Broad cell compatibility | Transfects a wide range of adherent and suspension cells and supports co-transfection of siRNA and plasmid DNA. | |
| High transfection efficiency | Efficient delivery of both plasmid DNA and RNA. | |
| Low cytotoxicity | Gentle on cells, with no need to change the medium after transfection. | |
| Serum compatible | Serum and antibiotics do not affect transfection, so complexes go straight into complete medium. |
Specifications
| Technology | Liposome-mediated (cationic lipid) transfection |
| Nucleic acids | Plasmid DNA and RNA (including siRNA); co-transfection supported |
| Cell types | Adherent and suspension eukaryotic cells, e.g. 293T, A549, HeLa, SW480, 4T1, Jurkat, NIH3T3, MDA-MB-453, Vero, K562, Raw264.7 |
| Recommended ratio | 1 μg DNA : 3 μl reagent (adjustable 1–5 μl per μg) |
| Medium | Compatible with serum and antibiotics; no medium change required |
| Shelf life | 2 years |
Components
| Components | Amount |
| GenieFect 2000 Transfection Reagent | 1.5 ml / 4 × 1.5 ml |
Storage
Store at 2 ~ 8°C.
Transient transfection in a 24-well plate is shown as an example.
- Seed the cells
- About 24 h before transfection, seed cells at about 0.5–2 × 105 cells/well.
- Incubate overnight.
- Form reagent/DNA complexes
- In a sterile 1.5 ml tube, mix 25 μl Opti-MEM with 1.5 μl GenieFect 2000 Transfection Reagent by gentle pipetting.
- In a second sterile 1.5 ml tube, mix 25 μl Opti-MEM with 0.5 μg DNA by gentle pipetting.
- Add the DNA/Opti-MEM to the reagent/Opti-MEM, mix gently by pipetting and leave at room temperature for 5 min.
- Transfect
- Add the complexes to the culture medium and gently rock the plate to distribute them evenly.
- Incubate for 24–48 h.
- If needed, replace with fresh serum-containing medium before transfection to prevent cell death from high density or nutrient depletion. If the medium must be changed after transfection, do so 6–12 h after adding the complexes.
Recommended starting conditions for other formats
| Culture vessel | 96-well plate | 24-well plate | 6-well plate | 6 cm dish | 10 cm dish |
| Surface area (cm2) | 0.35 | 1.9 | 9.6 | 20 | 59 |
| Serum-free medium (μl) | 2 × 5 | 2 × 25 | 2 × 100 | 2 × 250 | 2 × 500 |
| GenieFect 2000 Transfection Reagent (μl) | 0.3 | 1.5 | 7.5 | 30 | 45 |
| Plasmid, 1 μg/μl (μl) | 0.1 | 0.5 | 2.5 | 10 | 15 |
| Complete growth medium (ml) | 0.1 | 0.5 | 2 | 5 | 10 |
These conditions are a starting point and can be optimised for your cells.
Q1: What reagent-to-DNA ratio should I use?
- For a new cell line, start with 3 μl reagent per 1 μg DNA (1:3). The reagent volume per 1 μg DNA can be adjusted between 1 and 5 μl to improve efficiency.
Q2: Do I need to remove serum or antibiotics, or change the medium?
- No. Serum and antibiotics do not affect transfection, so complexes can be added directly to complete medium and no medium change is required. If you do change the medium after transfection, wait 6–12 h after adding the complexes.
Q3: What cell density should I transfect at?
- Generally 60%–80% confluence, but the optimum varies by cell line, so confirm it in a pilot experiment. Keep seeding ratios consistent for reproducible results.
