At first I used for my transformations by electroporation CL990-cin in concentration 100 mkg/ml. But single colonies of not-transformed strain grow on a plate. I increased concentration of antibiotic to 300, but it didn't help. A substitution of CL990-cin to Zeocin is also. Plates were made without direct light source and covered by foil. It only leaves pH. But may be someone had any experience with this antibiotic?
Hi,
Usually 100ug/ml.
However certain yeast strain can be resistant So you can test you host with different concentrations.
Regards
Hi,
As Nokuthula suggests, usually, peoples use 100ug/ml zeocin concentration for P. Pastori's culture.
The high concentration of Zeocin could help in the transformed clone screening. You can use 100-300ug/ml of Zeocin. 200ug/ml worked for me.
To see the colonies on Zeocin plate, the proper transformation protocol should be followed and consider the factors affecting transformation efficiency like
Way the competent cell prepared
nature of the recovery medium
Cell density
DNA concentration
Best
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