GSM8226335: Whole Bone Marrow, Mouse 919, NPM1cA/SMC3+/-, Rep1; Mus musculus; RNA-Seq
Myeloid sarcoma tumors were collected en masse and dissected away from surrounding tissue and directly placed in Trizole reagent with total RNA isolation and polyA mRNA isolation using NEBNext Poly(A) mRNA magnetic isolation module (NEB, E7490); Whole bone marrow was collected from tibias, femurs and pelvic bones by crushing in PBS buffer (containing 1% pen/strep, 0.5%FBS, and 0.5mM EDTA). Filtered through 70uM filter then spun down and red blood cells were lysed with Red Blood Cell Lysis Buffer, washed with additional PBS buffer, strained through 40uM filter and spun down. Cells were resuspended in PBS buffer and placed in Trizole reagent. Total RNA isolated and polyA mRNA isolation using NEBNext Poly(A) mRNA magnetic isolation module (NEB, E7490). Libraries were constructed using polyA mRNA magnetic isolation module (NEB, E7490), ERCC RNA Spike-In Mix (ThermoFisher Scientific, 4456840) was added, followed by NEBNext Ultra II RNA library prep kit for Illumina (NEB, E7770), the NEBNext multiplex oligos for Illumina set 1 and set 2 (NEB, E7335 and E7500) and Ampure XP Magnetic Beads (Beckman Coulter, A6388). Libraries quantified using NEBNext lirary quant kit for Illumina (NEB, E7630). PhiX control v3 (Illumina, 15017666)