What contributes to successful transfection?

Cell line health and viability, nucleic acid quality, transfection reagents, duration of transfection, and serum requirements all affect the success of transfection. Cells must be healthy, uncontaminated, and in the correct growth phase and density (typically 40-80% confluent for chemical methods). Nucleic acid quality (high purity, low endotoxin levels) and appropriate reagent concentration are also crucial for optimal outcomes.

Is it possible to predict the transfectability of a specific type of cell?

No, predicting transfection efficiency based solely on cell type is not possible. The interactions between cell properties and transfection reagents are complex. However, it is empirically known that some cell types, such as primary cells, quiescent cells, and suspension cells, are generally difficult to transfect.

What impact does the type and quality of DNA or RNA have on transfection success?

High-purity genetic material consistently leads to better transfection outcomes. Endotoxin contamination, introduced during bacterial processing, must be removed as it can trigger the innate immune system and hinder transfection. Additionally, promoter characteristics, gene properties affecting cell physiology or survival, and the nucleic acid's size and structure all influence transfection success.

How is Transfection Different from Transformation and Transduction?

Transfection refers to the introduction of foreign nucleic acids into cells, typically by non-viral methods, especially in animal cells. Transformation describes the same process in plant, fungal, and bacterial cells, but is avoided in animal cell biology due to its association with 'malignant transformation' in cancer. Transduction refers to the viral-mediated transfer of nucleic acids into cells.

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