GSM5589199: SUM159 CTR HI rep2; Homo sapiens; RNA-Seq
Cells extracted from tumor masses were washed in PBS 1% BSA and pellets were resuspended in blocking buffer (PBS 10% BSA) and incubated for 30 min at 4C light protected. After a wash in PBS 1% BSA, cells were incubated  for 45min at 4C, with FITC-conjugated anti-human CD24 antibody (Miltenyi Cat. #: 130-112-844) and vioblue-conjugated anti-human CD44 antibody (Miltenyi Cat. #: 130-113-899) staining. Finally, cells were washed with PBS 1% BSA and sorted with MoFlo Astrios Cell Sorter (Beckman Coulter).  Total RNA was isolated from cells, previously sorted, using the miRNeasy Mini Kit (QIAGEN, #217004), according to the manufacturer's instructions  mRNA-seq indexed library preparation was performed starting from 500 ng of total RNA with the TruSeq stranded mRNA (Illumina).