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Limited dilution of S2


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#1 christinam

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Posted 01 October 2009 - 11:02 AM

I have a stable cell line that produces a low yield of my protein so I'm trying to clone the cells by limited dilution. I have the cells in 15% heat inactivated FBS 100%conditioned media with hygro but my cells aren't dividing and it's been a week. Any suggestions?

#2 bob1

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Posted 01 October 2009 - 03:26 PM

Did you titrate a kill curve for the hygromycin before you used it?

Are the cells senescent?

#3 christinam

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Posted 02 October 2009 - 08:50 AM

My hygro is very low50yg/ml. at the low end of a kill curve I made a year ago one another clone. Many cells are dying but many more just seem stopped. I did this exact procedure 2y/o and it worked perfectly.




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