Hi all,
I was wondering if anyone used Zeocin to select positivie recombinant in continuous or intermitent light for more than 5 days? any thoughts about how stable it will be, knowing that it is sensitive to light exposure...Do you think I should continuously add Zeocin in my medium in case it got degraded/ destabilized?
Thanks in advance,
This may useful for you
https://www.researchgate.net/post/Is_Zeocin_a_good_antibiotic_selection_for_Ecoli_transformations
The Zeocin appears to have very mild selectivity, for example transformed cells grow only slightly faster than untrans formed cells on Zeocin agar plates
It worked for 6 weeks in low light at 18 degrees with a long day photoperiod (16h Light/8h Dark)
Hi Fatima,
I have used Zeocin in continuous light for transformation and selection of diatoms. From the start of selection it generally takes nearly 12 days. By day 14 of selection big colonies appear. Keeping the plates beyond 14 days increases appearance of false positives. This must be also balanced with the initial plating density, if too many cells are plated at the beginning of selection, it takes longer for transgenic colonies to appear and also there are too many escape (non-transgenic) colonies. For diatoms we use sea salts based medium and I have observed that best selection is obtained at 1/2 to 1/4 salt strength. This is all based on agar plates, although it can be used for maintaining selection in liquid cultures as well.
Hope this helps.
Best,
Yogesh
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