GSM5831855: Total RNA Eu-Tcl1 mouse 1; Mus musculus; RNA-Seq
Healthy and CLL splenic B cells were isolated from one-year-old WT and Eμ–TCL1 mice by   positive B-cell selection with CD19 magnetic beads (CD19 MicroBeads, Miltenyi Biotec, cat. 130-121-301). Briefly, spleens were squashed, treated with red blood lysis buffer (homemade)  and filtered in PBS using 40-mm cell strainers. The cells were then counted and the B cells were purified according to the manufacturer's protocol. Splenocytes subjected for polysome profiling were isolated using buffers (above) supplied with 100 ug/ml Cycloheximide (Sigma-Aldrich) to form 80S-mRNA complex. Five ug of RNA extracted from WT and Eµ-Tcl1 purified B cells together with RNA extracted from the three main polysomal fractions (Free, Light and Heavy) were subjected to library preparation of Cap Analysis Gene Expression CAGE using CAGE-seq protocol adapted for Illumina sequencing (Takahashi, Hazuki, et al. "5′ end–centered expression profiling using cap-analysis gene expression and next-generation sequencing." Nature protocols 7.3 (2012): 542-561.)