( ite cell line will either die out ...)Transfection Reagent Selector Kit Handbook

ite cell line will either die out or acquire a stable, heritable alteration, giving rise to a continuous cell line. This alteration is commonly known as in vitro transformation or immortalization, and frequently correlates with tumorigenicity.

Transfection Considerations

Media and supplements

Media are composed of a mixture of essential salts, nutrients, and buffering agents. Sterile media are usually purchased in solution. Alternatively, packaged premixed powders are available. Most media purchased are guaranteed to be mycoplasma- and endotoxin-free. Supplements to the media must include glutamine and can include nonessential amino acids, sodium pyruvate, and antibiotics. Some common media include DMEM, F12, DMEM/F12, RPMI 1640, MEM, and S-MEM.

Serum

In most cases media are supplemented shortly before use with serum. Fetal calf serum (FCS) is often used, but for some applications less expensive sera like horse- or calf serum can be used. Generally serum is a partially undefined material, that contains growthand attachment factors and may show considerable variation in the ability to support growth of particular cells. Variations in the serum quality can also lead to variation in transfection efficiency. In general, it is advisable to test a small lot of serum from a reputable supplier with a control cell line and assay before performing transfection experiments. Once a given lot has been shown to yield satisfactory and reproducible results, additional sera from the same lot should be purchased.

Transfection methods

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