GeticoFect Multi Transfection Reagent
Product Ordering Information
Catalog Number | Specification | Price (CNY) |
181901 | 0.75 mL | 3,800.00 |
181902 | 1.5 mL | 6,280.00 |
181903 | 15 mL | 56,520.00
|
1. Introduction
The GeticoFect Multi Transfection Reagent, developed by Getico Scientific (Shanghai) Co., Ltd., is a versatile and high - performance transfection reagent. It is designed to facilitate the efficient delivery of various nucleic acids, including DNA (plasmids), RNA, siRNA, mRNA and miRNA, into a wide range of eukaryotic cells, both adherent and suspension. This reagent enables researchers to perform a variety of experiments, such as gene expression studies, protein production, gene silencing, and functional genomics research.
2. Kit Components
- GeticoFect Multi Transfection Reagent: The key component of the kit, formulated to form stable complexes with nucleic acids for efficient cellular uptake.
- Positive Control DNA (GFP - Encoding Plasmid): Included to verify the transfection efficiency. This plasmid encodes for green fluorescent protein (GFP), allowing for easy visualization of transfected cells.
- Transfection Enhancer (Optional): Some kits may contain a transfection enhancer that can be used to further boost transfection efficiency. It works in synergy with the transfection reagent to promote better uptake of the nucleic acid - reagent complexes by the cells.
3. Storage Conditions
- Store the GeticoFect Multi Transfection Kit at 4°C. Do not freeze any of the components. Freezing can lead to precipitation, aggregation, and inactivation of the reagents, which will significantly reduce the kit's performance.
- When stored under the recommended conditions, the kit has a shelf life of 12 months from the date of manufacture.
- Avoid repeated exposure to room temperature. Return the kit components to 4°C immediately after use.
4. Pre - transfection Preparation
4.1 Cell Culture
- Cell Line Selection and Maintenance: The reagent is compatible with a wide range of eukaryotic cell lines. Ensure that the cells are in a healthy, actively growing state before transfection. Different cell lines may have specific culture requirements. For example, common cell lines like 293T, HeLa, and CHO cells can be cultured in appropriate media such as DMEM, RPMI - 1640, or Ham's F12K, usually supplemented with 10% fetal bovine serum (FBS) and 1% penicillin - streptomycin. Passage the cells regularly, splitting them at an appropriate ratio (e.g., 1:3 to 1:6) when they reach 80 - 90% confluence.
- Cell Seeding: For adherent cell lines, seed the cells in the appropriate culture vessels (e.g., 24 - well plates, 6 - well plates, 60 - mm dishes) the day before transfection. Aim for a cell confluence of 60 - 70% on the day of transfection. For example, in a 24 - well plate, seed approximately 1 - 2 x 10⁵ adherent cells per well. For suspension cell lines, adjust the cell density to 1 - 2 x 10⁶ cells/ml in the culture vessel on the day of transfection.
4.2 Nucleic Acid Preparation
- Type and Quality: The reagent can be used with different types of nucleic acids. For DNA, ensure high purity, with an A₂₆₀/A₂₈₀ ratio between 1.8 - 2.0. Plasmid DNA should be free of endotoxins, as endotoxins can interfere with the transfection process and cause cytotoxicity. For RNA, use RNase - free techniques to prepare and handle the RNA to prevent degradation. siRNA and miRNA should also be of high quality and stored properly at - 20°C or - 80°C until use.
- Quantity: The optimal amount of nucleic acid for transfection depends on the type of nucleic acid, the cell line, and the experimental purpose. As a general guideline, for plasmid DNA transfection in a 24 - well plate, start with 0.5 - 2 μg of DNA per well. For siRNA and miRNA, a concentration range of 50 - 200 nM can be used. It is recommended to perform a titration experiment to determine the optimal amount for your specific experiment.
5. Transfection Procedure
5.1 Complex Formation
- Reagent Equilibration: Take out the GeticoFect Multi Transfection Reagent and, if applicable, the transfection enhancer from the refrigerator and allow them to equilibrate to room temperature for 10 - 15 minutes.
- Dilution of Reagent and Nucleic Acid:
- In a sterile tube, dilute the nucleic acid in serum - free medium (such as Opti - MEM).
- In a separate sterile tube, dilute the GeticoFect Multi Transfection Reagent in the same serum - free medium. The ratio of the transfection reagent to nucleic acid needs to be optimized. As a starting point, for plasmid DNA transfection, a ratio of 1 - 3 μl of transfection reagent per 1 μg of DNA can be used. For siRNA and miRNA, a ratio of 1 - 2 μl of transfection reagent per 50 - 100 nM of nucleic acid can be used.
- If the kit contains a transfection enhancer, add the appropriate volume of the enhancer to the tube containing the diluted transfection reagent and mix gently.
- Then, add the diluted nucleic acid to the tube with the transfection reagent - enhancer mixture and mix gently by pipetting up and down 3 - 5 times.
- Incubation: Incubate the mixture at room temperature for 15 - 30 minutes to allow the formation of stable nucleic acid - reagent complexes. During this time, the reagents and nucleic acid will interact to form complexes that are ready for cell uptake.
5.2 Transfection
- For Adherent Cells: After the complex formation, add the nucleic acid - reagent complexes directly to the wells containing the adherent cells. Gently swirl the culture plate to ensure even distribution of the complexes. There is no need to remove the culture medium containing serum before transfection, as the reagent is compatible with serum - containing media.
- For Suspension Cells: For suspension cells, transfer the appropriate volume of cell suspension into a sterile tube. Then, add the pre - formed nucleic acid - reagent complexes to the cell suspension and mix gently by pipetting or gentle vortexing at a low speed. Transfer the transfected cell suspension back to the culture vessel.
5.3 Post - transfection Incubation
- Cell Incubation: Incubate the transfected cells in a humidified incubator at 37°C with 5 - 8% CO₂. For short - term studies (e.g., observing transfection efficiency by fluorescence microscopy for a reporter gene), results can be observed as early as 24 - 48 hours post - transfection. For long - term studies (such as recombinant protein production or gene silencing over several days), the cells may need to be incubated for an extended period. Monitor the cell growth and viability regularly during the incubation period.
6. Optimization
- Reagent - Nucleic Acid Ratio: Since different cell lines and nucleic acid types may respond differently to the transfection reagent, it is essential to optimize the ratio of the GeticoFect Multi Transfection Reagent to nucleic acid. Perform a series of experiments with varying ratios while keeping the amount of nucleic acid constant (e.g., 1 μg of DNA) and changing the volume of the reagent (e.g., 0.5 μl, 1 μl, 1.5 μl, 2 μl, 2.5 μl, 3 μl) to determine the ratio that gives the highest transfection efficiency with the lowest cytotoxicity.
- Nucleic Acid Concentration: In addition to optimizing the reagent - nucleic acid ratio, the concentration of the nucleic acid itself may need to be optimized. Test different concentrations of nucleic acid (e.g., for DNA, 0.25 μg, 0.5 μg, 1 μg, 1.5 μg, 2 μg per well in a 24 - well plate) while maintaining the optimal reagent - nucleic acid ratio to find the best conditions for your specific experiment.
- Cell - Specific Optimization: Some cell lines may have unique characteristics that require additional optimization. For example, certain sub - lines may be more sensitive to the transfection conditions, or they may respond better to transfection at a specific cell density. Adjust the cell seeding density, incubation time, and other parameters as needed to improve transfection efficiency.
7. Troubleshooting
7.1 Low Transfection Efficiency
- Check Nucleic Acid Quality: Ensure that the nucleic acid is of high purity and integrity. Degraded or contaminated nucleic acid can lead to low transfection efficiency. Re - purify the nucleic acid if necessary.
- Optimize Reagent - Nucleic Acid Ratio: As mentioned above, re - optimize the ratio of the reagent to the nucleic acid. Try different ratios and concentrations to find the optimal conditions for your cell line and nucleic acid type.
- Cell Health and Density: Check the health of the cells. Unhealthy or over - confluent cells may have reduced transfection efficiency. Passage the cells at the appropriate time and ensure they are in a good physiological state before transfection. Adjust the cell seeding density if necessary.
- Complex Formation: Ensure that the nucleic acid - reagent complexes are formed correctly. Follow the incubation time and mixing procedures precisely. If the complexes are not formed properly, they may not be efficiently taken up by the cells.
7.2 High Cytotoxicity
- Reduce Reagent Concentration: If high cytotoxicity is observed, try reducing the amount of GeticoFect Multi Transfection Reagent used in the transfection. Lower the reagent - nucleic acid ratio and see if the cytotoxicity decreases while still maintaining an acceptable transfection efficiency.
- Check Incubation Time: Prolonged incubation time may increase cytotoxicity. Shorten the incubation time of the transfection complexes with the cells, for example, from the conventional overnight incubation to 4 - 6 hours, then replace with fresh medium to observe if the cytotoxicity is reduced.
- Cell Handling: Ensure that the cells are handled gently during the transfection process. Rough handling, such as excessive pipetting or vigorous shaking, can damage the cells and increase cytotoxicity.
8. Safety Precautions
- Personal Protective Equipment: Wear appropriate personal protective equipment, including lab coats, gloves, and safety glasses, when handling the kit components.
- Avoid Ingestion and Inhalation: Do not ingest or inhale any of the reagents in the kit. Keep the kit components away from the mouth, nose, and eyes. In case of accidental contact with the eyes, rinse immediately with plenty of water and seek medical advice.
- Chemical Storage: Store the kit in a secure location away from heat, light, and incompatible chemicals. Follow the storage conditions mentioned above to maintain the stability and integrity of the reagents.
- Disposal: Dispose of any unused kit components and waste materials according to local regulations. The reagents may be considered chemical waste and should be handled appropriately.
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