4 protocols
AccessionType
sample treatment protocol
5-Azacytidine treatment at final concentration of 5μM for 24 hrs (at 32 hrs post infection by lentivral vectors)
nucleic acid extraction protocol
After the last washing round the protein-RNA adducts were subjected to RNA fragmentation on the beads, using RNA fragmentation reagent (Ambion) at 94C for exactly 5min in a thermocycler, followed by 1min incubation on ice and then quenched by fragmentation stop solution (Ambion). The fragmented RNA was then purified by ethanol precipitation followed by Trizol (Invitrogen) RNA extraction and used for library preparation. Illumina’s directional mRNA-Seq sample preparation protocol
sample treatment protocol
5-Azacytidine treatment at final concentration of 3μM for 12 hrs (at 18 hrs post infection by lentivral vectors)
nucleic acid extraction protocol
Total RNA was purified from an aliquot of 5-Azacytidine treated cell lysate, before performing the immunoprecipation (IP). PolyA mRNA was isolated using oligo(dT) beads followed by mRNA fragmentation and library preparation according to Illumina's "directional mRNA-Seq sample preparation protocol".