EzFect DNA transfection reagent
Non-liposomal formulations exhibit low toxicity, and the transfection efficiency in commonly used tool cells reaches 90%.
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Instruction manual- Product Description
- Composition
- Precautions
-
Product preservation
Store at 4°C; shelf life is 12 months. For long-term storage, keep at −20°C. Transport at room temperature.
Product Overview
This product is a next-generation, water-soluble, highly charged cationic polymer–based transfection reagent. It forms stable complexes with negatively charged plasmid DNA, which then adhere to the negatively charged cell surface and are internalized through the cell membrane. This reagent is suitable for the efficient, transient transfection of plasmid DNA into adherent or suspension‑cultured cells and functions optimally in serum‑ and antibiotic‑containing complete culture media. Even when transfecting plasmids as large as 13 kb, this reagent maintains high transfection efficiency.
Product Features
1. High transfection efficiency, simple operation, low toxicity, and excellent reproducibility. For most cell lines, such as HEK293T, HEK293F, Expi293, CHO, and HeLa, transfection efficiency reaches 80%–90% when the DNA-to-transfection reagent ratio is maintained at 1:1 to 1:2 (based on DNA amount in μg and reagent volume in μL).
2. Excellent compatibility: transfection can be performed in the presence of serum and antibiotics, without the need to pre‑replace Opti‑MEM® or other serum‑free media.
3. The transfection reagent is a chemically synthesized product and contains no animal-derived components (Animal origin-free, AOF).
4. This product has been sterilized by filtration through a 0.22 µm membrane filter.
Instructions for Use (DNA Transfection)
(This instruction uses a 6-well plate and 293T cells as an example.)
1. Cell seeding
The day before transfection, seed 293T cells in a 6-well plate at 3 × 10^5 to 5 × 10^5 cells per well, and culture them in DMEM supplemented with 10% serum for 18–22 hours.
2. Transfection Process
a) Prepare DNA dilution (per well): Add 2 µg of plasmid DNA to 100 µL of serum-free DMEM, vortex for 5 seconds, or pipette up and down 20 times.
b) Preparation of the transfection complex (per well): Add 3 μL of transfection reagent directly to the DNA dilution, vortex for 5 seconds or pipette up and down 20 times, and let stand at room temperature for 20 minutes (do not exceed 30 minutes).
c) Use a pipette to dispense the transfection complex into the cell culture wells, and gently rock the 6-well plate to mix thoroughly.
d) Transfect for 10–16 hours, then remove the old culture medium and replace it with 2 mL of fresh DMEM containing 10% serum.
e) Continue culturing for 24 to 48 hours, then proceed with the subsequent experiments.
Reference Table for DNA and Transfection Reagent Dosages Across Different Cell Culture Vessel Sizes
Cell culture vessel Culture medium volume Volume of the diluted solution DNA amount Transfection reagent 96-well plate 100 µL 10 µL 0.1 µg 0.15 µL 48-well plate 200 µL 20 µL 0.2 µg 0.3 µL 24-well plate 500 µL 50 µL 0.5 µg 0.75 µL 12-well plate 1 ml 50 µL 1 µg 1.5 µL 6-well plate 2 ml 100 µL 2 µg 3 µL 6 cm Petri dish 4 ml 200 µL 4 µg 6 µL 10 cm Petri dish 10 ml 500 µL 10 µg 15 µL Instructions for Use (siRNA Transfection)
(This instruction uses a 6-well plate and a final siRNA concentration of 50 nM as an example.)
1. Cell seeding
The day before transfection, seed an appropriate number of cells in a 6-well plate to achieve a cell density of 30%–70% at the time of transfection.
2. Transfection Process
a) Take a sterile 1.5 mL centrifuge tube, add 100 μL of serum-free culture medium first, then add 100 pmol of siRNA, and mix well.
b) Add an additional 5 μL of transfection reagent to the siRNA, immediately vortex for 5 seconds or pipette up and down 20 times, and incubate at room temperature for 20 minutes (do not exceed 30 minutes).
c) Use a pipette to dispense the transfection complex into the cell culture wells, and gently rock the 6-well plate to mix.
d) Transfect for 10–16 hours, then remove the old culture medium and replace it with 2 mL of fresh complete culture medium.
e) Continue culturing for 24 to 48 hours, and assess the expression level of the target gene using qPCR or Western blot.
Reference table for siRNA and transfection reagent amounts based on cell culture vessel size
Cell culture vessel Culture medium volume Volume of serum-free culture medium for dilution siRNA dosage Transfection reagent 24-well plate 500 µL 50 µL 25 pmoles 1.25 µL 12-well plate 1 ml 50 µL 50 pmoles 2.5 µL 6-well plate 2 ml 100 µL 100 pmoles 5 µL 6 cm Petri dish 4 ml 200 µL 200 pmoles 10 µL 10 cm Petri dish 10 ml 500 µL 500 pmoles 20 µL -
Product Composition
Item number Component Specifications 100-505 ezFect DNA transfection reagent 1 mL -
Precautions
1. Prior to transfection, cells must be in good growth condition. Using cells that are in the logarithmic growth phase, exhibit high viability, and have been passaged only a few times will yield better transfection results.
2. The quality of plasmid DNA is critical for successful transfection. It is recommended to use high‑purity, endotoxin‑free plasmids. The amount of plasmid DNA should be optimized according to the cell type and the specific experimental conditions.
3. Depending on the specific experimental conditions and cell type, the DNA (µg)/transfection reagent (µL) ratio is recommended to be optimized within the range of 1:1 to 1:4.
4. This product is intended solely for scientific research by qualified professionals and shall not be used for clinical diagnosis or treatment, nor for food or pharmaceutical purposes, nor stored in ordinary residential premises.
5. For your safety and health, please wear a lab coat and disposable gloves while performing this procedure.
6. If you encounter any issues during use, please contact our technical support.
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