title: NextSeq 500 paired end sequencing; RNA-seq of human MDS/AML cell line OCI-M2 treated with 5-azacytidine and sulforaphane
design: RNA-seq of human MDS/AML cell line OCI-M2 treated with 5-azacytidine and sulforaphane
library_construction_protocol: OCI-M2 AZA-R subclones evolved from AZA-S cells after treatment with AZA doses exceeding IC50 for 8 weeks. AZA-S and AZA-R cells were incubated with 1 µM 5-azacytidine, 5 µM sulforaphane or the combination of both for 24h. Total RNA from freshly harvested cells were isolated with Trizol reagent following manufacturers protocol. 1µg of DNase I-treated RNA isolated from treated cells was used for poly-A mRNA isolation with the NEBNext Poly(A) mRNA Magnetic Isolation Module. Paired-end libraries were prepared using the NEBNext Ultra II Directional RNA Library Kit with NEBNext multiplex index oligos for Illumina (Dual Index Primers Set 1) (New England Biolabs).