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?
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#2
bob1
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Thelymitra pulchella
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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?
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.
