title: GSM2399087: cKO2; Mus musculus; RNA-Seq
library_construction_protocol: Before FACS staining, DN thymocytes were highly purified by depletion of CD4+ and CD8+ and DP thymocytes with AutoMACS. Then, DN NKT cells were FACS sorted using BD FACS Aria, and RNA was prepared using RNeasy Micro Plus kit containing gDNA eliminator columns (Qiagen). RNA was analyzed for quantity and quality using Agilent Bioanalyzer 2100 or Agilent 2200 TapeStation equipment, and 10 ng of total RNA was used with SMARTer Ultra Low Input RNA Kit v3 for Illumina Sequencing (Clontech) for cDNA synthesis. Libraries were prepared according to NEB's instructions accompanying the NEBNext® ChIP-seq Library Prep Reagent set for Illumina.  Adapter ligated DNA was PCR amplified with Illumina primers for 15 cycles. The purified DNA was captured on an Illumina flow cell for cluster generation. Libraries were sequenced on the Hi-Seq 1500 following the manufacturer's protocols.
identifiers.namespace: GEO
identifiers.id: GSM2399087