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G418 selection problem - (May/22/2006 )

I have selected my transfected cells with G418 for 10 days. But there are still some untransfected control cells attached to dish. Unlike healthy cells, these cells are just round-shaped. Are these cells dead or not? Should I increase G418 concentration?

-macrosky-

round shaped means they don't attach strongly. But not means 100% dead. Don't increase the G418 selection, but maintain pressure for additionnal week and you'll probably get rid of untransfected cells.

-fred_33-

Recently I selected my NIH3T3 cells using G418 (200 ug/ml). It took me about a month to get nice colonies to pick up. So have pacience - the mock will eventually die. Give it a month or so (depending on your line)

-liflaf1-

Thank you guys. Then I will wait and wait.

QUOTE (liflaf1 @ May 23 2006, 02:14 AM)
Recently I selected my NIH3T3 cells using G418 (200 ug/ml). It took me about a month to get nice colonies to pick up. So have pacience - the mock will eventually die. Give it a month or so (depending on your line)

-macrosky-

has anybody done the stable selection work in mES cells?

QUOTE (macrosky @ May 24 2006, 04:20 AM)
Thank you guys. Then I will wait and wait.

QUOTE (liflaf1 @ May 23 2006, 02:14 AM)

Recently I selected my NIH3T3 cells using G418 (200 ug/ml). It took me about a month to get nice colonies to pick up. So have pacience - the mock will eventually die. Give it a month or so (depending on your line)

-JerryLee-