title: Direct RNA-Seq of SLU7-AA Saccharomyces cerevisiae: RNAs treated with rSAP, E.Coli PolyA Polymerase and Terminator Exonuclease (BAM)
design: Saccharomyces cerevisiae cells were grown in YPD media for 2 doublings before being shifted to media containing 1ug/ml rapamycin for 1 hour. Total RNA was extracted via phenol chloroform extraction. In order for hmsnoRNAs to be sequenced with direct RNA sequencing the samples were first treated with Shrimp Alkaline Phosphatase in order to remove 3' phosphates from snoRNAs. Then samples were in-vitro polyadenylated using recombinant E. Coli polyA polymerase. Then RNAs were treated with Terminator 5' to 3' exonuclease to remove ribosomal RNAs. Libraries were then prepared using Oxford Nanopore Direct RNA sequencing kit according to manufacturer's instructions.