Biomedical Engineering Reference
In-Depth Information
produce multiple glycerol stocks of each verified bait and prey clone. In addition, avoid
perpetuating clones on selective plates without rechecking auto-activation patterns before
screening.
•
When using the PJ69-4A strain in combination with either pGBAD-B/pACTBD-B-,
pGBAE-B/pACTBE-B- or pACT-based clones; we have occasionally observed 'inter-
actions' which strongly activate the ADE2 reporter, but not the HIS3 or lacZ reporters.
These interactions appear to be false positives. This phenotype emphasizes the need for
using multiple reporters to score Y2H data.
•
The original manuscript describing the PJ69-4A yeast strain suggested that the use of the
liquid-nitrogen snap freezing procedure when performing
β
-Gal assays could be prob-
lematic, leading to auto-activation of the lacZ reporter gene. In our experience using the
pGBAD-B or pACTBD-B series of vectors, we have not found this to be the case.
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