GSM9267380: mRNA_WT_for_tif3_a; Saccharomyces cerevisiae; RNA-Seq
Cells were rapidly vacuum filtered and flash frozen in liquid nitrogen. Whole cell extracts were prepared by breaking cells in freezer mill in cell lysis buffer. Total mRNA isolated from whole cell extract using miRNeasy minikt (Qiagen). Total RNA fragmentation, Size selection, linker ligation, reverse transcription, cDNA circularization, PCR amplification) was done as described Ingolia NT, Ghaemmaghami S, Newman JR, Weissman JS. 2009. Ribo-Zero Gold rRNA Removal Kit (Illumina) was used to to remove ribosome contamination after linker ligation step.