title: GSM3593783: Ime4 KO Yeast PolyA RNA Direct RNA Sequencing rep1; Saccharomyces cerevisiae SK1; RNA-Seq
library_construction_protocol: Yeast total RNA samples were prepared using a modified protocol of nucleospin® 50 RNA kit (Machery-Nagel, cat 740955.50). Specifically, cells lysis was done in a 1.5ml tube by adding 450µl of lysis buffer containing 1M sorbitol (SIGMA-ALDRICH), 100mM EDTA and 0.45µl lyticase (10U/µl). The sample was incubated in 30°C for 30 minutes to break the cell wall, centrifuged for 10' at 3000 rpm, and supernatant was removed. From this stage, extraction proceeded as in the protocol of nucleospin® 50 RNA kit, only substituting β-mercaptoethanol with DTT. Enrichment of polyadenylated RNA from total RNA was performed using Oligo(dT) dynabeads mRNA-DIRECT kit (Thermo Scientific, 61012) for small mRNA amounts.  RNA libraries were prepared by following the standart ONT direct RNA sequencing (SQK-RNA001) protocol. Model: GridION R9.4.1 Flowcell FLO-MIN106 MiKNOW 1.15.1
attributes.tag: GEO Accession
attributes.value: GSM3593783