Antibiotic selection Puromycin versus Neomycin


This was posted by someone else a couple years ago (no replies), but I thought I would try again, perhaps more people are using this technique now??

I would like to try antibiotic resistance method for selecting LARGE numbers of transgenic worms over several/many generations in liquid culture ---- Using either puromycin (Nature Methods, 2010) or using neomycin (Nature Methods, 2010). Does anyone use either of these methods and have any advise or opinion on which antibiotic selection has more advantages over the other and what are the limitations found by you. (or sources for the antibiotic selection marker vectors for microinjection? )

Cheers muchly :slight_smile:

Hi Kitska10,

We’ve used both selection markers in the Jorgensen lab. Personally, I prefer the neomycin (G418) selection. Neo is quite a bit cheaper than puromycin and works very well in our hands. I haven’t used it in liquid cultures but Neo works quite well by top-spreading already seeded plates (we add 500 ul of a 25mg/ml stock solution dissolved in water to regular “small” plates and let them dry down on a benchtop). That way you don’t have to plan very far ahead. The drug does not kill adult worms but will arrest/kill any progeny hatched on the plates. So as long as you keep the cultures growing vigorously I would expect it to work quite well.

Both the Dupuy and Lehner labs distribute the antibiotic resistance plasmids via Addgene. And both labs have been very helpful when I needed advice.

Good luck,

Thanks heaps for that Christian, and you are getting very pure transgenic populations?
I’ll look into getting the vector.