Instrument: Illumina HiSeq 1000
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: SINGLE
Construction protocol: CD34 positive cord blood cells were isolated using magnetic beads from fresh cord blood (For each biological replicate, three different cord blood samples were mixed). The cell were retrovirally infected with ETV6-NCOA2 or empty vector and cultured in IMDM supplemented with hSCF, hTPO, hFlt3L (100 ng/ml) for 5 days. The positive cells were sorted (GFP). We extracted RNA from the samples and performed bulk RNA-sequencing. RNA was purified, analyzed by bioanalyzer (Agilent 2100), cDNA libraries were prepared using Using PrepX mRNA Library kit (WaferGen), and Apollo 324 NGS automatic library prep system. Using the universal (SR) and index-specific primer with limited PCR cycle number (~13), a sample-specific index was added to each adaptor-ligated cDNA sample. Libraries were clustered onto Illumina's TruSeq SR Cluster kit v3, and sequenced for 50 cycles using TruSeq SBS kit on Illumina HiSeq system