GSM5129420: DETC Tcrd-/- A; Mus musculus; RNA-Seq
The desired populations were sorted and the RNA was isolated employing the "RNeasy Plus Micro kit" from Qiagen (Hilden, Germany). The cDNA was generated by employing ingredients from the TAKARA cDNA Synthesis Kit (TAKARA Bio Europe, France) according to an in-house modified protocol based on the manufacturer's protocol. The DNA-amplicons for the Illumina sequencing were generated by using Advantage 2 PCR Kit (TAKARA Bio Europe, France). The DNA was then purified using Qiaquick Gel Extraction Kit (Qiagen Hilden, Germany). The purified product of each sample was indexed by an indexing-PCR with Nextera-Primers. Finally a pool of the samples was sequenced employing Illumina MiSeq v2 Reagent Kit (Illumina Inc, San Diego, CA, USA).