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refrozen cell pellet

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#1 scramble



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Posted 10 March 2011 - 10:29 AM

I grew a few liters of large culture (e.coli cells) to purify some membrane proteins. I harvested them and froze them at -80C. I took them out a few days later, thawed them, and resuspended them in buffer containing protease inhibitors, thinking that I was gonna lyse and do a purification that day... However, I did not need the cells... So I spun the cells back down, dumped out the buffer, and put them back in -80C. Is the cell pellet not good to use anymore?? Or is this usually ok?

#2 protolder



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Posted 13 March 2011 - 10:42 PM

Hola,The only thing that you have get is to have some of your bacteria broken , so the first stage to lyse is done, but you have to sonicate or DNAse digest, to have the DNA broken and avoid the viscosity of the sample. Buena suerte

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