Peptide-based nucleic acid delivery is a state-of-the-art delivery technology that can significantly improve nucleic acid transfection of a wide range of cells, including primary cells and hard-to-transfect cell lines. Based on the high-throughput peptide screening platform, GentleFectTM technology accurately identifies endogenous peptide sequences with phase separation potential, and developed the Gentle A30 nucleic acid transfection reagent for hard-to-transfect cell lines. Peptides are self-assembled by liquid-liquid phase separation with nucleic acids to form nano-scale droplets, which can be efficiently internalized with the help of cell membrane pinocytosis, resulting significantly improvement of the nucleic acid delivery efficiency of cells across all species while cell viability are also maintained.
• The product is suitable for transfection of various nucleic acid molecules, including DNA, mRNA, siRNA, circRNA, Crispr, etc.;
• The product is a ready-to use transfection reagent, easily add nucleic acids to the reagent to configure the transfection solution in a single step.
4°C for short storage and -20°C for long storage.
• Gentle A30 Nucleic Acid Transfection Reagents for Hard-to-Transfect Cells: Reagent A, Reagent B
Suspension cells are cultured in suspension, allowing for a higher cell density per well compared to adherent cells. Both types of cells can be transfected during cell passaging, so adherent cells do not need to be plated before use. In addition, with numbers of experiments tested, transfect cell suspension mixture in an EP tube help maximize the transfection efficiency.
It is recommended to follow the transfection reagent amounts specified in Instruction manual . Alternatively, the amount of transfection reagent can be adjusted proportionally according to the number of cells, also suspension cells can be transfected according to the common density used in customer’s cell culture system.
① Optimize Cell Condition: Prior to transfection, ensure cells viability exceed 90%. This can be assessed using standard viability assays such as trypan blue exclusion or propidium iodide staining.
② Adjust Reagent Concentrations: Incrementally increase the amounts of transfection reagent and nucleic acid to identify the optimal ratio that maximizes transfection efficiency without compromising cell viability.
③ Optimize Incubation Time: Experiment with varying incubation times to determine the ideal duration for your specific cell type. Both prolonged and shortened incubation periods can influence transfection efficiency and cell viability, so it is essential to find a balance that yields the best results.
① Do not use medium containing FBS during cell transfection, FBS will significantly compromise the efficiency of cell transfection;
② Gentle A30 Transfection Reagent is not recommended to vortex vigorously, and it is recommended to use a pipette to mix by gentle and repeated pipetting.
① The centrifugation protocols described in this manual are standardized at room temperature, 300g, for 5 minutes.
② After transfection, add 1 mL of complete medium and mix gently to terminate the transfection process and mitigate the potential impact of the transfection reagent on subsequent cell centrifugation (Note: The slight viscosity of the transfection reagent is typical and does not indicate an issue);
③ After centrifugation, the cell pellet may not be readily visible, especially when working with low cell numbers. This is a common occurrence and does not necessarily indicate a procedural issue.
Gentle A30 Nucleic Acid Transfection Reagent For Hard-to-Transfect Cells v1.3
