An effective method is the freezing and thawing of the algal biomass.

Thanks monica

Hm tried sonication? Or alcohol bath?

Sonication can be very effective, but it depends on the structure of the cell wall and membrane. This paper describes and details the effectiveness of a number of methods (and whether subcellular components remain intact/functional).

Michel Lavoie, Jonathan Bernier, Claude Fortin, and Peter G.C. Campbell (2009).

"Cell homogenization and subcellular fractionation in two

phytoplanktonic algae: implications for the assessment of metal

subcellular distributions" Limnology and Oceanography : Methods. p. 277 - 286.

thanks jacqueline . i have tried sonication but i could not separate my protein of interest when doing SDS PAGE..i will go through the paper and try again.

Disruption using a French press can also be useful. In this process, cells in liquid medium are placed in a cylinder and subjected to high pressure, then a small orifice is opened. Cells break as they depressurize while passing through the orifice. Using a French press will avoid some of the heating that can occur in sonication.

Ultimately, no single method is ideal for all samples--be prepared to experiment. Good luck with your project.

Thanks eric .and i want to ask that woud u specify any particular instrument for this procedure? It will be more useful to me.

The French Press I used was from Thermo Scientific, but I think the product is now unavailable. However, a similar model appears to be available from Glen Mills (http://www.glenmills.com/cell-disruption/high-pressure-homogenizer/french-press-gm.html).

There are a number of reviews on cell disruption methods (not limited to algae), e.g., http://www.google.com/url?sa=t&rct=j&q=&esrc=s&source=web&cd=1&cad=rja&ved=0CCYQFjAA&url=http%3A%2F%2Ftur-www1.massey.ac.nz%2F~ychisti%2FCellDRev.pdf&ei=zRluUNrIK6Lq2QXCloDIAQ&usg=AFQjCNHntFg-ivdb_gkSTavN2rjSQjHVww.

Combination of freeeze thawing and soniccation is the very effectivemethod for rupturing algal biomass.mainly for protein seperation technique.

Soniccation followed by freeze thawing and again soniccation

Why rupture? You can identify and quantify algae composition in-vivo using NMR. You only need about 1ml. The main drawback is that sensitivity is not so high (about 1% , but better if you know what you are looking for). The main advantage of NMR is that your material is preserved and can be reused for breeding colonies with the selected characteristics.

If you send me an email ( j.defeijter(at)hts-110.com ) with some information about your research I may be able to give you some more suggestions.

I've got best results with freezing the pelleted algal biomass with liquid nitrogen with subsequent grinding it in frozen state (i.e. without thawing) and immediate extraction of the ground sample.

You can use 50mM Tris-HCL treatment for 6 hour.

Hi @Vidya: I think the main problem with you the preparation of buffer. Please prepare buffer in a good optimal pH range. Otherwise if you use anyone of the methods the results should appear on SDS-PAGE. For Qualitative assay, all the methods will produce results. Check your buffer preparation again. And one more thing, what happens in lab that all the students use same chemicals so they may contaminate one with other. Try prepared buffer solution from some company.

The best way to rupture the algae cell walls is to use a Microfluidizer, which is a high pressure fluid processor that provides high shear rate to help rupture wide variety of cells. Technologies such as french press and freeze-thawing are not reproducible, ultrasonication usually generates very high local temperatures near the probe which results low yields. Besides, they all have the scalability issues. You can learn more about the Microfluidics' technology and its benefits at

http://www.microfluidicscorp.com/applications/biotechnology

Here are a few references on cell disruption:

http://www.ncbi.nlm.nih.gov/pubmed/22946699

http://www.ncbi.nlm.nih.gov/pubmed/25268351

http://www.ncbi.nlm.nih.gov/pubmed/22690952