how to separate membrane from cytoplasms in diatom

[eldesiempre]

Hi folks,

 

I have some diatoms in culture and I am looking for literature about lysis and subsequent separation of cell components (membrane from cytoplasmatic constituents). I was wondering if anyone here has worked on that topic, or even if you have any tips.

 

cheers

[CharonY]

One simple way is ultracentrifugation after lysis. For more fractions creating a suitable density gradient can be used. The latter is a little bit tricky.

[Greippi]

Oddly enough, I did this yesterday.

 

But I did it on bacterial cells, maybe it's different for diatoms.

 

I French pressed my cells to lyse them. Then I applied the stuff to the top of a sucrose gradient. Ultracentrifuge over night, then harvested the membrane band.

 

I can give you a more detailed protocol if you like - but the conditions will probably be different for diatoms. It also depends on what you intend to do with the membranes.