Cell culture is a particularly tricky endeavor. The media, environmental conditions, and technique need to be just right to ensure that the experiments are reproducible and dependable. For this type of research, you will need the appropriate selection antibiotics to verify your gene of interest is incorporated into your cell line. You may also need antimycotics to control fungi that could arise. Lastly, you may require Mycoplasma detection and elimination products since this type of contamination can cause cause alterations of various cell characteristics which may lead to inaccurate experiment conclusions. A wide range of antimicrobials are used in transfection protocols to ensure the gene is incorporated, and many others are used to control contamination from invading microorganisms.
Transfection is the process of introducing foreign nucleic acids (like DNA or RNA) into a eukaryotic cells using non-viral methods. This allows researchers to study gene function and protein expression. There are two main types of transfection: a) Transient transfection (the nucleic acid is temporary) and b) Stable transfection (the nucleic acid integrates into the cell genome). Transfection is done with chemical methods (like using lipids or polymers) or physical methods (like electroporation or microinjection). If you are doing transfection, our popular video series walks you through the steps involved using G418 Disulfate and chinese hamster ovary (CHO) cells as an example.
Plant transfection, also known as plant transformation is done using biological methods involving the bacterium Agrobacterium tumefaciens. Other methods are physical methods like biolistics (gene gun) and electroporation (via plant protoplasts). Other techniques involve treatment with chemicals or sonoporation. All of these techniques can introduce foreign nucleic acids into plant cells for research, and gene selection agents are used.
If you are starting a new cell culture project, consult our website or digital flyer to get started. Our Cell Culture Database is also a great resource to review which antimicrobials you should use for your cell line. It can help you determine the concentrations and combinations of antibiotics for gene selection and transfection, looking for published studies for your cell line of interest.
If you are interested in saving some time in the media prep lab, our popular ReadyMade® antimicrobial solutions arrive sterile-filtered and ready to use which means one less antimicrobial stock solution to prepare. With the right tools and techniques at your disposal, your experimental cell lines can be healthy, clean, and ready for their close-up.