返回
返回
返回
Eukaryotic Cell Transfection Reagent Manual: Instructions for Use of a High-Efficiency Transfection Reagent
EZ Trans cell transfection reagent, a next-generation transfection agent, operates on the principle that a positively charged cationic polymer forms a positively charged complex with the negatively charged phosphate groups of nucleic acids. This complex then interacts with the negatively charged proteoglycans on the cell surface and is internalized via endocytosis. Optimized by Li Ji Bio, this product boasts high transfection efficiency, low cytotoxicity, ease of use, excellent reproducibility, and broad applicability.
The following is the instruction manual for eukaryotic cell transfection reagents:
Transfection Reagent Transportation and Storage
Blue ice transport. Store at 4°C; shelf life: 12 months. [Note]: Do not freeze!
Instructions for Using the High-Efficiency Transfection Reagent (Using a 24-well plate as an example; for transfection in other culture dishes, please refer to Table 1)
1. Cell seeding
For adherent cells, plate them 18–24 hours prior to transfection (without antibiotics) to achieve a confluence of approximately 80% at the time of transfection. For suspension cells, plate them on the day of transfection before preparing the EZ Trans–DNA complex, adding 4–8 × 10^5 cells per 500 µL of growth medium.
[Note]: Serum in the culture medium does not affect transfection efficiency. The amount of transfection reagent required depends on the cell type and other experimental conditions; it is recommended to perform a dose‑response titration when first using the reagent to optimize the optimal dosage.
2. Prepare the EZ Trans-DNA complex
(1) For each well, dilute 1 μg of plasmid DNA in 40 μL of serum-free high-glucose DMEM (or OPTI-MEM I medium), and mix thoroughly.
(2) For each well, dilute 3 μL of EZ Trans transfection reagent in 40 μL of serum-free high-glucose DMEM (or OPTI-MEM I medium), and gently mix.
[Note]: Serum-free high-glucose DMEM is the diluent; serum-containing media must not be used to dilute DNA or EZ Trans transfection reagents, as the formation of the EZ Trans–DNA transfection complex requires a serum-free environment.
(3) Add the diluted EZ Trans transfection reagent to the diluted plasmid DNA as quickly as possible, and gently mix.
[Note]: The order of this mixture must not be reversed.
(4) Incubate at room temperature for 10–15 minutes to form the EZ Trans-DNA complex.
3. Transfected cells
(1) Pipette the aforementioned 80 μL of EZ Trans‑DNA transfection complex evenly into the culture dish containing the cells. Gently rock the dish or apply mild shaking to ensure uniform dispersion of the EZ Trans‑DNA complex.
(2) Incubate in a 37°C, 5% CO₂ incubator for 6–18 hours; remove the culture medium containing the EZ Trans–DNA complex, replace it with fresh medium, and continue culturing until analysis of the transfected gene expression.
[Note]: To select stably transfected cell lines, after transfection, passage the cells into fresh growth medium (diluting the cells at least 10-fold), incubate at 37°C in a 5% CO₂ incubator for 24 hours, and then add selection medium. Under selection conditions with a drug that corresponds to the transfection‑resistant gene, drug‑resistant clones can typically be isolated within approximately 1–2 weeks; during this period, the growth medium containing the selection agent should be replaced frequently.
Precautions
-
This product is intended solely for scientific research and must not be used for clinical diagnosis, treatment, or any other purposes.
Group Website
-
Contact Information
Landline phone:021-50778506
Business Manager:18616108315Business Cooperation/Scholarships:18616108697
Address:4th Floor, Building 27, Lane 908, Ziping Road, International Medical Park, Pudong New Area, Shanghai
Email:ztt0005@obiosh.com
WeChat Official Account
Online Customer Service 1
Online Customer Service 2
Telephone consultation
