GSM9042119: eIF3g/i; Saccharomyces cerevisiae; RNA-Seq
SELEX was performed for 12 rounds in each experiment, as described  in 5, with 18 base-long randomized ribonucleotide sequence flanked by primer binding sites designed for cDNA synthesis, PCR and RNA synthesis by T7 RNA polymerase. For these experiments we used His6-tagged eIF3g/Tif35p or its complex with (untagged) eIF3i/Tif34p. The DNA fragments encoding them were amplified by PCR using budding yeast Saccharomyces cerevisiae genomic DNA and then cloned into pET28b (with hexa-histidine tag fused to its N-terminus, in the case of eIF3g) and were co-expressed in E. coli strain BL21-CodonPlus (DE3)-RILP (Stratagene).  The expressed protein(s) was purified by Ni-NTA (Qiagen) and Q-sepharose (Cytiva). SELEX output RNAs were ligated to adaptor sequences and made into cDNA, which was then sequenced by generating RNAseq library