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Stable Cell Selection with Neomycin


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4 replies to this topic

#1 Gam8Doc

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Posted 11 June 2009 - 07:36 AM

My plasmid that I am using has the neomycin resistance gene and we are using two breast cancer cell lines, MDA-MB 231 and PMC42LA. When I did my kill curve I had to use concentrations of 6mg/ml to get any cell death. This seems really high and I wanted to make sure this was on par with other peoples experiences with Neo. Also at this high of a concentration it turns my media yellow, so I want to make sure that it is truly killing my cells by the antibiotic, not some changes to the media

Thanks

#2 bob1

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Posted 11 June 2009 - 05:01 PM

200-400 ug/ml is the usual range, try these concentrations and leave for a little longer, it should work fine.

#3 Gam8Doc

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Posted 12 June 2009 - 11:45 AM

This was for 12 days and my 1mg/ml plate was growing as well as the control.

#4 Vini

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Posted 13 June 2009 - 11:11 PM

200-400 ug/ml is the usual range, try these concentrations and leave for a little longer, it should work fine.



i normally use a concentration of 100 ug/ml for my cell culture. at max. i go upto 200. while increasing the concentration gradually i make sure that cells are growing in an expected way.

#5 pcrman

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Posted 14 June 2009 - 08:17 AM

You got such high resistance concentration (6mg/ml) which is not real. For antibiotics selection, one important issue is to seed the cells as single cells at low density. When cells are confluent, they become very resistant.




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