GSM3777049: AML7 high; Homo sapiens; RNA-Seq
The mononuclear cell (MNC) fraction from BM and/or PB was obtained by density gradient centrifugation using lymphoprep (PAA, Cölbe, Germany) according to standard procedures. For separation of ROSlow and ROShigh fractions following thawing of viably frozen MNCs, cells were washed and after overnight recovery on MS5 cells were stained for CD34+ and CellROX. Subsequently cells were sorted for CD34+, 15% low and 15% high ROS fraction. After sorting cells were lysed in TriZol for RNA extraction. RNA libraries were prepared for sequencing using standard Illumina protocols