Hi Eleanor,
Does blank = yeast cells without the lacZ gene? If it is a control without
cells, how long do you incubate after adding ONPG? Old ONPG solutions can
hydrolyze spontaneously, so it is a good idea to make them fresh. If you
are incubating for long times, try increasing the concentration of your
cells instead; that may reduce the colour that you are seeing in your
control.
David
Hi,
I am working with the liquid beta-gal assay with ONPG as substrate. I'm also
using SDS+chloroform to destabilize cell walls. I seem to have high beta-gal
activity in my blank (as yellow color development). Has anyone observed the
same. I'd be grateful for any ideas especially as I'm not a microbiologist.
Thanks, Eleanor.
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