RNA binding of Saccharomyces cerevisiae Ssd1

We quantified the exact RNA binding sites of the Ssd1 protein in Saccharomyces cerevisiae, in exponential growth and heat shock conditions, using the CRAC protocol. We used a His-TEV-protein A tag (HTP) on the C-terminal of the genomic copy of Ssd1, with the 3'UTR replaced by the 3'UTR/terminator from the K. lactis Ssd1 homolog, followed by a KlURA3 selection marker. Overall design: Five samples: two biological replicates of CRAC binding to Ssd1-HTP tagged protein at both of 30C and 42C 16min, and a negative control sample of background strain BY4741 with no tagged protein.