title: GSM4222244: WT_4705_B; Saccharomyces cerevisiae; RNA-Seq
library_construction_protocol: Newly synthesized RNAs were labeled as previously described (Bonnet et al., 2014; PMID: 25228644). S. cerevisiae and previously labeled S. pombe cells were mixed in an 8:1 ratio and total RNA was extracted using the RiboPure yeast kit (Ambion, Life Technologies) using the following volumes:  480 μl lysis buffer, 48 μl 10% SDS, 480 μl phenol:CHCl3:IAA (25:24:1)  per S. cerevisiae pellet + 50 μl S. pombe (from a single S. pombe pellet resuspended in 850 μl lysis buffer). Total RNA was then biotinylated and purified essentially as described (Duffy et al., 2015; PMID: 26340425; Duffy and Simon, 2016; PMID: 27925666) using 40 μl (~40 μg) total RNA. In the last step 50 μl each of RNA sample eluted from streptavidin beads was adjusted to 100 μl with nuclease-free water and purified on RNeasy columns (Qiagen) using the modified protocol as described (Duffy and Simon, 2016; PMID: 27925666). RNAs were eluted into 14 μl nuclease-free water. Ovation Universal RNA-seq System kits (Tecan)
attributes.tag: GEO Accession
attributes.value: GSM4222244