Several labs have developed antibiotic selection markers that are very useful for transgenesis (see below). We’ve modified the selection protocols for efficient selection for miniMos insertions. The large advantage of the antibiotic selection markers is that: 1) they can be used in any genetic background and 2) insertions can be followed in subsequent crosses by adding antibiotics to plates. A disadvantage of the antibiotic selection markers is that it is harder to identify homozygous insertions in the absence of relatively bright fluorescence because non-resistant worms are hard to identify on plates with antibiotics.<\/p>\n
We find that adding 500 ul of an antibiotic stock solution to NGM plates already seeded with bacteria is easier than pouring NGM plates with antibiotics and this protocol takes a lot of the planning out of experiments. In our hands the selection is still very efficient and equivalent to unc-119<\/em> selection. The selection works best on growing populations of worms and not very well on starved populations. Adult worms without the resistance transgene generally survive several days so injected worms can be placed directly onto plates with the antibiotic selection or the antibiotic can be added 1-2 days after injection. In our hands, Neomycin (G418) and hygromycin selection works faster and is slightly more efficient compared to puromycin selection.<\/p>\n\n