GSM2418344: RNAseq DN2 WT 1; Mus musculus; RNA-Seq
Harvested cells were stained with lineage marker, CD25, CD117, ghost dye red 780. Lineage negative DN2 cells were sorted by FACS AriaII and washed with PBS. Total RNA was isolated using RNeasy Mini kit (Qiagen). RNA was treated with TURBO DNase (Life Technologies). mRNA was purified from total RNA using Dynabeads mRNA purification kit (Life Technologies). cDNA was generated with the First-strand synthesis kit (Life Technologies) using a random hexamers in presence of actinomycin D. Second-strand synthesis was performed with dUTP instead of dTTP. Sonicated cDNA was ligated to adaptors (NEBNext primer kit). The resulting DNA was treated with uracil-N-glycolsylase prior to PCR amplification with the indexing primers (NEBNext primer kit).
GEO Accession
GSM2418344