2 replies to this topic

[BioBus]

HI,
after sonication,  and centrifuge with top speed for 30 min
at  4 degree,
there is  a layer of lipids floating on the supernatant..


How you treat them?  

Certainly I cannot ignore them , but no idea how they
produced and how to further  proceed.

They seems  definitively among  the useless for  IP .

thx,

Bus

[KPDE]

BioBus, on Jul 22 2009, 06:20 AM, said:

HI,
after sonication,  and centrifuge with top speed for 30 min
at  4 degree,
there is  a layer of lipids floating on the supernatant..


How you treat them?  

Certainly I cannot ignore them , but no idea how they
produced and how to further  proceed.

They seems  definitively among  the useless for  IP .

thx,

Bus

I usually just ignore the small pellicle that I get after centrifugation.  If you think it might be a problem then you could always increase the ratio of sonication buffer to cells.  Of course this means you have to sonicate a larger volume so, if you do this, you might want to split your lysate into two aliquots before sonicating.

[bob1]

Just remove the solution from under the lipid layer to a new tube.  Or, conversely take the lipid layer off the tube and discard, it shouldn't contain anything of importance for your IP.