Hi,
I have a serious problem and hope so that someone of you
could help me:
Almost all of my plates (80 !) of my yeast two hybrid
screening (GAL 4 System, Y190) seem to be contaminated with bacteria!
The plates are now incubating for 6 days and several yeast cultures
(approx. 0.5 mm diameter)have formed all over the plates, as expected.
But all over the plates (in the area around the colonies)
a light "smear" is visible.
I streaked over a part of some plates and streaked the cells
on a slide.
Under the microscope many healthy yeast are visible as well
as some yeast with clear bacteria IN it (they are moving in the yeasts
cells) as well as single bacteria cells and some clusters of bacteria
forming around lysed yeast cells.
After that,I picked single yeast colonies, they are looking healthy and
hardly unaffected.
What can I do now?
Do I have to throw all these 80 (!) plates away and start a new
screening?
Or could I still use them?
Will the bacteria seriously affected the results of the screening?
Could I somehow "spray" any antibiotic on the plates?
Normally, I would like to incubate the plates for another four days
before performing the beta-Galactosidase-Assay.
Should I wait these days or should I proceed as fast as possible?
I would imagine that the E.coli might have overtaken the
complete plates and killed all yeasts if I will wait longer.
Do you have experiences with that?
Thank you so much in advance for your answer!
Everything would help!
Thorsten
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