GSM7727715: MPL-Calr(del52) Bone Marrow 5; Mus musculus; RNA-Seq
Bone marrow cells were isolated from femur, tibia and hip bone on day 21 past BMT. Erythrocytes were removed using erythrocyte lysis buffer. Cells were strained through a 75 µm cell strainer to receive single cell suspension. For all steps cells were treated according to 10xGenomics  Cell preparation guidelines. Samples were processed using the Chromium Next GEM Single Cell 3' Reagent Kit v3.1 (Dual Index – 10x Genomics) following manufactures protocol. In short: GEM´s were generated using a Chromium Controller (10x Genomics) at a target cell recovery rate of 10.000. GEM-RT and subsequent clean-up was performed. cDNA was amplified and 3' Gene Expression Dual Index Library Construction was performed. Illumina paired-end, dual indexing was performed. For all amplification steps a Veriti 96 Well Thermal Cycler (appledbiosystems) and for all quality control steps a 2200 TapeStation (Agilent Technologies) was used.