Which drug-selection marker should I use?
VectorBuilder offers a variety of drug-selection markers on its vectors, such as puromycin (Puro), neomycin (Neo), hygromycin B (Hygro) and blasticidin (Bsd). In general, we have found that puromycin kills non-resistant cells faster and more consistently than other antibiotics. For this reason, we recommend Puro for most cell types. However, some cell types may naturally have some degree of resistance to certain antibiotics without the resistance gene or that they are sensitive to certain antibiotics even with the resistance gene. These cells would require testing of different drug-selection markers to find the optimal ones.
Recommended concentration and selection duration of commonly used drugs
| Antibiotics | Cell line | Recommended concentration | Recommended duration |
|---|---|---|---|
| Puromycin | 293T | 1-2 ug/ml | 3-5 days |
| Geneticin (G418) | HT1080 | 500-1000 ug/ml | 7-11 days |
| Blasticidin | 293T | 5-15 ug/ml | 7-11 days |
| Hygromycin B | 293T | 100-200 ug/ml | 5-7 days |
Our VectorAcademy post Enhance Your Culture: How to Improve Bacterial Growth provides a guide to optimizing antibiotic concentration as well as other culture conditions to enhance plasmid yield.
Note:
a. Geneticin (G418) is used to select for cells expressing neomycin resistance gene.
b. Passaging cells frequently may speed up blasticidin selection.
Read more about our collection of drug-selection markers