A pool of 5,896 haploid-convertible heterozygous diploid yeast knockout (YKO) strains were transformed with a cdc102D::natMX cassette. The resultant diploid double mutant pool was again transformed with a cdc102-1 allele on a centromere-based plasmid, which also carries a URA3 gene as the selection marker. This cdc102-1 pool was sporulated and the spores were spread on MM-Ura (as target source 1) and MM-Ura+Nat (as target source 2). Pools of haploid YKOs were harvested after incubation at 28 degree for 2 days. Genomic DNA samples were prepared from these two haploid pools and used as the templates for PCR amplification of the TAGs identifying the YKOs with a pair of primers universal for either Uptags or Downtags. The PCR products from sample1 were labeled with Cy5 and those from sample2 with Cy3. The Cy5- and Cy3-labeled Uptags and Downtags were mixed together and hybridized to a Hopkins-TagArray. Fluorescent array images were collected for both Cy5 and Cy3 with a GenePix4000B microarray scanner. The image data was annalyzed with a GenePix5.1 program. Keywords = cdc102-1, 28 degree, heterozygotes, YKO
