what procedure we will follow if we have to isolate the cell wall of green algae from rest of the components?

Question

techniques , procedures and references required for PhD research

Answer 1

You could use sucrose-step gradient centrifugation. This is where, in a centrifuge tube, a density gradient is created. This means that at the bottom of the tube, there will be a layer of say 70% sucrose, and on top of that, a layer of say 50% and so on. This will vary depending on the sample you want to look at. You would 'mash up' the algae and place the sample on top of the sucrose layers. Then you would spin at a very high speed, around 150000 x g. The cell wall particles will be pulled through the different layers until they come to the one with the same density. You can then isolate this fragment. (btw, sucrose is good for neutral, water soluble molecules). You can do a search for sucrose step gradient centrifugation and there are a wealth of papers which will give details on how the technique is carried out.