Yeast Protein Protein Interactions Protocols

Genome-Wide Analysis of Protein-Protein Interactions Using a Two-Hybrid Array This procedure capitalizes on the asexual/sexual reproductive cycle of yeast cells. Laboratory yeast strains can be maintained indefinitely by asexual reproduction as one of two haploid mating types, MATa or MAT{alpha}.

Genome-Wide Analysis of Protein-Protein Interactions Using a Two-Hybrid Array: This protocol describes how to handle arrayed yeast using robotics.

Genome-Wide Analysis of Protein-Protein Interactions Using a Two-Hybrid Array: Amplification of ORFs This protocol describes the first step in constructing an array: amplification of the predicted ORFs that are to be included in the array. Gene-specific primers containing vector-specific flanking sequences that facilitate recombinational cloning are used to amplify each ORF. A secondary amplification can be used to extend the length of the homologous vector sequence flanking the ORF.

Genome-Wide Analysis of Protein-Protein Interactions Using a Two-Hybrid Array: S This protocol describes three different storage methods for the array. Frozen stocks are the most permanent method and are an absolute requirement for maintaining a record of the array.