96-well Beta-gal assay for yeast ?
bjorn.johansson at tmb.lth.se
Sat Oct 10 11:17:50 EST 1998
Dear Yeast Community,
I would like to ask a question regarding microtiter plate Beta-Gal
assays for yeast. I know this issue has been discussed here before,
but I haven't found any concluding answer.
I would like to grow my yeast cells in a 96-well plate, permeabilize
them and assay for Beta-galactosidase with ONPG directly in the plate.
I have read the article of F. Kippert 1995, FEMS Microbiology letters
"A rapid permeablization procedure for accurate quantitative
determination of Beta-Gal. in yeast". There is a method described using
Sarcosyl and DTE and no need for vortexing. I wonder if any one has
done this in 96-well plates ?
Some formulas for calculating Beta-gal activity from permeabilized
cells involve subtracting OD550 from OD420 to make up for cells in the
cuvette. Is it possible to leave out the centrifugation step and
measure OD420 and OD550 on the cellsuspension and still get a good
activity reading ? I don't have a centrifuge for microtiter plates, so
I would prefer not to have to spin down the cells.
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