NextSeq 500 sequencing; Perturbation of mRNP biogenesis reveals a dynamic landscape of the Rrp6-dependent surveillance machinery trafficking along the yeast genome (RNA-seq dataset)
Perturbation of mRNP biogenesis reveals a dynamic landscape of the Rrp6-dependent surveillance machinery trafficking along the yeast genome (RNA-seq dataset)
S. cerevisiae cells were grown according to standard procedures at 25 °C in synthetic complete medium with bases and amino acids omitted as necessary for selection and with glucose (2%) as a carbon source. The pCM185 vector (CEN, TRP1) from Euroscarf was used as backbone to create the Doxycycline-regulated (Tet off) Rho expression construct (pCM185-Rho-NLS) (Mosrin-Huaman et al., 2009). Strains transformed by Rho expression vector were grown under repressing conditions (5 µg/mL of Doxycycline) at 25°C in YNB medium supplemented with SC-TRP (Synthetic Complete Supplement Mixture from MP Biomedicals) and glucose as carbon source (final concentration 2%). To allow the expression of Rho, the yeast cells were grown the same medium but without Doxycycline. Growth was performed during 16 hours incubation at 25°C.  Total RNA extraction was performed using the acidic hot phenol/chloroform method  Libraries preparations were made by I2BC high-throughput sequencing platform (Gif sur Yvette) using Illumina ScriptSeq protocol after RiboZero treatment.