MYTH Screening: iMYTH and tMYTH Variants

Once a bait has been generated and validated for the membrane yeast two-hybrid (MYTH) assay, it can be used for either high-throughput screening to generate a detailed interaction map (interactome) or in low-throughput experiments to examine interactions with specific targets. Here we describe how to carry out high-throughput MYTH library screening of a validated bait generated using integrated or traditional MYTH. The principles herein can be easily adapted for use in a smaller-scale format if required. A typical MYTH library screen can be completed in ∼3-4 wk.