Tips for Transfection
- NanoEntek
- Sep 12
- 2 min read
Overview
Transfection is a core technique is the process of deliberately introducing foreign genetic materials (like DNA, RNA) into eukaryotic cells. For a detailed introduction to transfection, click HERE.
Even when working with the same cell type, transfection efficiency and results can vary depending on experimental conditions. This post focuses on the key factors that influence transfection efficiency and provides practical tips to maintain high cell viability after transfection.
Major factors affecting the transfection efficiency
✓ Cell condition
Cell viability is one of the most critical factors for successful transfection. When viability decreases, transfection efficiency also lowers, as confirmed by multiple studies. So, always use healthy, well-maintained cells.
✓ Cell culture media without antibiotics
Avoid using antibiotics in the culture media when plating cells for transfection. This is especially important for physical transfection methods like electroporation, where cells experience temporary damage and need recovery time. The presence of antibiotics prevents cells from healing properly, leading to reduced viability and lower transfection efficiency.
✓ Optimal quantity and quality of transfection agents
The quantity of transfection agent will highly affect the transfection efficiency. Too much may lead to cytotoxicity, whereas too little may not be sufficient to achieve successful transfection. Also, the quality of transfection agents is significant to getting acceptable transfection results.
✓ Appropriate cell density
Cell confluency strongly influences transfection efficiency because overcrowded cells can block nucleic acid entry while sparse cells may not proliferate enough for meaningful results.
✓ Short passage numbers
Keeping short passages for cell lines in consideration of the transitions in characteristics of cell lines over a certain number of passages is important. These changes often lead to inconsistent transfection results. To minimize variability, work with cells at lower passage numbers whenever possible.
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