GSM2454147: RNA Tbx6+ KO TS14; Mus musculus; RNA-Seq
Total RNA from approximately 20,000 cells was isolated using the RNeasy Micro kit (Qiagen) and genomic DNA was digested on column with the addition of an extra 1ul of RNase-free DNase I (Roche) to ensure complete digestion. The RNA from approximately 20,000 cellswas quantified using the Qubit RNA HS Assay and the integrity was verified using Bioanalyzer RNA Pico chips. Approximately 80-100ng of total RNA was used for the generation of strand-specific RNA-Seq libraries using the ScriptSeq v2 (Epicentre) low input libarary preparation kit according to manufacturer’s instructions. During library preparation, the purification after rRNA-depletion was performed using RNeasy Micro columns and the cDNA was purified using the MinElute PCR Purification Kit (Qiagen). The library was finally amplified with 15 PCR cycles.