Kurtiss observed as I finished wrapping up a Yeast two-hybrid experiment. I had already ran through the entire protocol, picked yeast colonies that grew on both double drop out and quadruple drop out agar plates with x-α-Gal (after mate and plate with a hcDNA yeast library), and streaked them out for isolation of a single yeast colonies. 15-cm agar plates were divided like a pizza and the sections were numbered. There were 9 colonies there were selected from plates, cultured for glycerol stocks and cultured for DNA purification. PCR was completed using designed primers and synthetic genes TRIM6, TRIM31 and TRIM69 for template.