Single and dual drug selection for transgenes following bombardment of Caenorhabditis species

• Drug selection markers provide a rapid, simple “hands off” selection method.
• As a dominant marker, drug selection can be used on any genetic background.
• Dual puromycin-G418 selection for bombardment works well in a variety of species.
• Single drug selection with either puromycin or G418 is also effective in C. elegans.
• Our liquid selection protocol is extremely economical and easy to parallelise.

The use of drugs and drug resistance genes is a powerful method to select for the presence of a transgene. Unlike methods that require the complementation of a genetic mutation, this system can be used on any genetic background. Drug selection does not require extensive manipulation or costly equipment, yet it is very rapid and can achieve extremely high efficiency, selecting a small number of transgenic worms from among millions of non-transgenic worms. Introducing integrated transgenes into Caenorhabditis elegans by microparticle bombardment represents just such a challenge. Here we describe in detail the protocol we have developed for dual-drug selection in liquid with puromycin and G418 which works well in a variety of Caenorhabditis species. We also show that single drug selection with only puromycin or only G418 is effective in C. elegans. The growing number of drug selection markers that have been adapted to C. elegans are an important addition to the genetic toolkit at our disposal.

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