title: GSM8891550: B-cells from young, Rps15 wildtype mouse, biological replicate 3 (Ribo-Seq); Mus musculus; OTHER
library_construction_protocol: Spleens or femurs of mice were mechanically dissociated into a single cell suspension. B-cells were negatively selected using the MACS Mouse B cell Isolation Kit (Miltenyi Biotec, #130-090-862). Total RNA was extracted with Qiagen RNAeasy kits. 200 ng per sample was used as input into an automated variant of the Illumina TruSeq Stranded mRNA Sample Preparation Kit (which preserves strand orientation of the RNA transcript). Oligo dT beads were used to enrich mRNA from the total RNA sample, followed by heat fragmentation and cDNA synthesis. The resultant 400 bp cDNA underwent dual-indexed library preparation, including 'A' base addition, adapter ligation using P7 adapters, and PCR enrichment using P5 adapters.