I have tried to use sodium alginate method to regenerate yeast
spheroplast. The methods is as follows:
1. Mix spheroplast with 2% sodium alginate.
2. Layer onto a petridish.
3. Add a layer of 0.4M CaCl2 to solidify the alginate.
4. Wash with KCl
5. Add medium to incuate.
The origninal reference is
Methods Enzymol. 1993;217:431-6.
Alginate as matrix support for yeast artificial chromosome cloning.
Cantrell C, Lai E.
I have not got any colonies from this method. Has anyone tried this
method? Anything I am not doing right?
Also, any other way to regenerate yeast spheroplast? I need at leat 50%
recovery for my use. Thanks!