Inducible meiotic cDNA libraries for genome-wide studies of gene isoforms

We report the construction of 5 yeast meiotic cDNA libraries and perform proof-of-principle screens to show that these yeast cDNA libraries can be used to identify genes and gene isoforms that are important for competitive fitness. Samples 1-25 are from different stages of cDNA library construction and deep sequencing was used to characterize gene representation in each yeast cDNA library. Samples 26-175 are from proof-of-principle competitive fitness screens. Synchronous meiotic timecourses were performed to obtain stage-specific mRNA, mRNA was converted to cDNA using TGIRT reverse transcriptase, Gateway cloning was used to insert cDNA libraries into galactose-inducible vectors, and plasmid libraries were transformed into yeast using electroporation. Competitive screens were performed in the indicated liquid media over 5 days.