title: GSM4494460: X35STRAPLATEOSROOT2; Oryza sativa Japonica Group; OTHER
library_construction_protocol: For Rice: For cellular mRNA, p35S:HF-FLAG-OsRPL8 pulverized tissue was extracted in PEB and a S16 fraction was obtained as detailed by Mustroph et al. 2009). Total poly(A)+ RNA was extracted by direct capture of poly(A)+ RNA using a biotinylated oligo dT according to Townsley et al., 2015. For TRAP mRNA, TRAP was performed as previously described (Mustroph et al 2009, Reynoso et al 2015) with the following modifications: ɑ-FLAG conjugated IgG Dynabeads were used for binding; after magnetic collection and washing the polysomes were removed from the magnetic beads by addition of Lysis and Binding Buffer (LBB) buffer for polyA mRNA isolation using biotinylated oligo-dT primers and streptavidin magnetic beads (NEB) (Townsley et al. 2014). RNA-seq libraries were prepared from poly(A)+ selected total and TRAP mRNA as described by Townsley et al. (2014) for random primer primed libraries.
identifiers.namespace: GEO
identifiers.id: GSM4494460