GSM7033888: B_strain_9307_6_D; Saccharomyces cerevisiae; RNA-Seq
RNA extraction was performed from yeast pellets using Acid Phenol:Chloroform pH 4.5 and Tris-EDTA-SDS (TES) buffer (0.01 M Tris-HCl pH 7.5, 0.01 M EDTA, 0.5% w/v SDS). Samples were further treated with rDNase (cat no 740.963, Macherey-Nagel) and column purified (cat no 740.948, Macherey-Nagel).  RNA levels were quantified by sequencing 3' RNA ends as previously described {Chapal, 2019 #326}. Briefly, RNA was reverse transcribed using oligo dT primers coupled with internal barcodes (Table S6). RNA-DNA hybrids were then pooled and tagmented using Tn5, adding adaptors to their 5' end, followed by PCR amplification of the fragments, inserting Illumina barcodes and adaptors. A total of four biological replicates were sequenced for each mutant-timepoint combination, two of each were processed in each of two pools. RNA-tagmentationSeq