Can anyone suggest a simple fractionation procedure for yeast suitable for
an undergraduate class? To cut down on time I was thinking of a density
gradient of Ficoll of maybe three concentrations to separate cytosol,
mitochondria, vacuoles and nuclei + debris (pellet?). Anyone tried this?
Also, I was thinking of using glass beads to rupture the yeast (cheaper
than lyticase!). This seems to work follwed by simple cntrifugation steps:
500, 12000 and 100000g), but a single density gradient step should be
quicker (and better?) in the end. This will be follwed by enzyme and
protein assays and western blotting of fractions (in subsequent classes).
Any advice gratefully received.
Fergus.Doherty at nottingham.ac.uk
0115 970 9366 (74-41366 internal)