I want to extract both bacteria and yeast but I can't find out best methods
I would recommend you to prepare your cell lysates using glass beads coupled to vortexing. I normally use glass beads of 500 µm diameter for yeast lysate preparation and it gives a good protein yield after purification.
There are many effective protocols but you can use this one from the European Molecular Biology Laboratory: http://www.embl.de/pepcore/pepcore_services/protein_purification/extraction_clarification/cell_lysates_yeast/glass_beads/
Finally, you have to remember to include proteinase inhibitors in your solutions in order to minimize any colateral damage during purification. I recommend to use a IP inhibitor cocktail kit EDTA free and also to subject to one freeze/thaw cycle at -80°C in order to weak yeast wall.
Make
Lysis buffer
50 mM Tris-HCl pH 8.0
150 mM NaCl
1 mM EDTA
1 mM PMSF
0.1 % Triton X
Pellet ur culture, add 4ml lysis buffer to (50 ml) pellet, vortex and keep in ice for two/four hrs, sonicate (10 sec off/on) for 5 min, pellet down (12000 rpm/ 20 min), keep the supernatant, this protocol works well with bacterial culture. Hope u will be benefited.
We used CelLytic B Cell Lysis Reagent from Sigma . According to Sigma, this is a highly efficient yet gentle reagent for the extraction of proteins from bacteria (E. coli). - See more at: http://www.sigmaaldrich.com/life-science/proteomics/recombinant-protein-expression/cell-lysis/bacterial-cell-lysis/cellytic-b.html#sthash.mcRuiNCM.dpuf
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