Defining a Functional Bait
Constructing and Transforming a cI Bait Protein
Constructing and Transforming a LexA Bait Protein
Replica Technique/Gridding Yeast: Assessing Bait Activation of Reporters
Detection of Bait Protein Expression
Combining the two Baits together and finalizing Baits characterization
Troubleshooting and alternative plasmids
Transforming and characterizing interactors from a library.
- Detailed library screening flow chart
Transforming the library
Harvest and pool primary transformants
Mating the Bait Strain and the Pretransformed Library
Mating the Bait and the Library Screening
First confirmation of positive interactions.
DNA isolation, and second confirmation of positive interactions.
PCR Approach_ Rapid screen for Interaction Trap Positives
Plasmid Isolation Approach: isolation of plasmids, transfer to bacteria
Followup for Library Screening