GSM1635423: Rag1ko ctrl-sh 3; Mus musculus; RNA-Seq
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 combination of random hexamers and oligo(dT) in presence of actinomycin D. Second-strand synthesis was performed with dUTP instead of dTTP. The ds-cDNA was sonicated to 200-400 bp using the S220 Focused-ultrasonicator (Covaris). 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). Following PCR, fragments were size-selected.
GEO Accession
GSM1635423