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Smokey stuff in 293T extract

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#1 AllenChiu



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Posted 30 May 2010 - 09:49 PM

Someone else and I have encountered with the smokey stuff in 293t extract for IP,initially they appear close to the surface of supernatant after 10min 12000rpm centrifuge of raw 293t lysis(with NP40 or TritonX100).
Someone in this forum said it was lipids,how could they know?and how to get rid them?Cause it seems sticky and I am afraid it will interfere the binding of beads.
In addition,after sonication the E. coli lysis will become super sticky,so why?because of DNA or lipids or?Will dilution be helpful?I sonicated E coli cell from 100ml of LB medium in 7ml of buffer,so was it too concentrated?what proportion between medium/cell paste and sonication buffer should be?

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