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3, broken yeast

Yeast is a generic name for a group of single-celled fungi, whose members belong to different fungi, cell wall composition will have a greater difference, these are to be done before breaking yeast cells to consider. I summed up a few ways to make broken yeast:

A mechanical method: the general PROTOCOL are used glass beads: such as sigma G-8772, plus glass beads after ultrasound, protein activity to maintain better.

2 methods of chemical lysis: 10g yeast plus 1ml of ethyl acetate, stir until liquid (hope expert review)

3 urea solution (urea 8mol / L, NaCl 0.5mol / L, Tris 20mmol / L, EDTA 20mmol / L, 2% SDS, pH 8.0) high pressure homogenization. (This seems to be the same in method 2)

4 liquid nitrogen freeze-thawed and ground yeast break, I think this may be appropriate in a small test.

5 Moderate enzymatic, may not destroy yeast protoplasts

Enzymatic cleavage mainly uses two kinds of enzymes: 1. Snail enzyme, can be obtained directly from the snail's gastric juice; lyticase, can be ordered from sigma. Both enzymatic methods can be used in combination with the above methods, especially the glass beads method, the effect is very good.

I used chemical lysis method, 10g yeast plus 1ml of ethyl acetate, stir until liquid. This method cracking good results, may wish to try.

An article is added urea lysis solution (urea 8mol / L, NaCl 0.5mol / L, Tris 20mmol / L, EDTA 20mmol / L, 2% SDS, PH value 8.0)

The method I used to make good yeast crushing was still glass beads, which, as microbe and rongjunli put it, are very efficient and have no effect on the activity of the protein of interest. Generally should use this method.