Strand specific RNA-Seq of Rituximab treated NSG-BL mouse tumors: Sample R0042
Strand Specific RNAseq Library Preparation and Sequencing. Total RNA was extracted from the eBL patient FNA samples, the resulting BL cell lines and NSG-BL mouse tumors using the Monarch Total RNA extraction kit (New England Biolabs). Starting with 1 to 5μg total RNA, we enriched mRNA molecules using oligo-dT magnetic beads to capture polyA-tailed mRNA molecules. We then proceeded to prepare strand-specific RNAseq libraries following the NEBNext® Ultra™ II Directional RNA Library Prep Kit for Illumina (NEB). Final library concentration and fragment size were confirmed with a qubit 4 fluorometer (Thermofisher) and fragment analyzer (Agilent) respectively. These libraries were sequenced with paired end reads (2 × 75 bp) on the NextSeq 550 system (Illumina, Inc.).