GSM2536331: mRNAseq_PADH1-GCN4S242L_met_rep2; Saccharomyces cerevisiae; RNA-Seq
A yeast cell pellet was resuspended in 1 ml of lysis buffer from Dynabeads® mRNA DIRECTTM Kit (61011, Life technologies) and lysed at 4°C with 1 volume of acid washed glass beads in a FastPrep instrument (Thermo Scientific) using 2 cycles with the following settings; 45 s at 6.5 speed with 3 min pause on ice between each cycle. Libraries for mRNA sequencing were prepared using the “directional ​mRNA­seq sample preparation” protocol from Illumina with minor modifications. Polysome profiling and sequencing of ribosome-protected mRNA fragments were performed according to protocol described in (Nedialkova and Leidel, 2015).