GSM3616160: Tumor_UTR5; Mus musculus; RNA-Seq
Single cell suspension of tumor and spleens was generated, CD11b+ cells were enriched by MACS sorting, live MDSC populations (PMN-MDSC= CD11b+Ly6CloLy6G+; M-MDSC =CD11b+Ly6ChiLy6G-) were isolated by FACS sorting. Bulk tumor samples were directly lysed in QIAzol Lysis buffer. RNA was extrated using the QIAzol Lysis buffer together with the Qiagen RNeasy Kit. Library for RNA sequencing was prepared using the SMARTer® Stranded Total RNA-Seq Kit v2 - Pico-Input Mammalian (TaKaRa). In brief, up to 10 ng RNA was fragmented for 4 min at 94°C, followed by first-strand cDNA synthesis. Next, Illumina Adapters and Indexes were added. Subsequently, RNA sequencing library was isolated using AMPure beads and ribosomal RNA was depleted using ZapRv2 and R-Probes v2. Finally, RNAseq library was amplified in 13 cycles und isolated using AMPure beads.
GEO Accession
GSM3616160