Direct RNA sequencing of in-vitro polyadenylated saccharomyces cerevisiae RNA

In order to extend the study of the accumulation of hybrid mRNA/snoRNA species in Saccharomyces cerevisiae for intron-hosted snoRNAs, libraries were sequenced using Oxford nanopore direct RNA sequencing. Libraries were prepared for WT control samples and Slu7 AA samples, which were shown to accumulate hybrid mRNA/snoRNAs. Overall sequencing strategy was to degrade rRNAs and non-pol II transcribed RNAs by phosphatase treatment from Shrimp Alkaline Phosphatase then digestion with Terminator 5-Phosphate dependent exonuclease. m7G capped RNAs would remain unaffected and in vitro polyadenylation would ensure that MinION compatible RNA adapters are able to be ligated to the remaining species.