GSM8449010: BM 4.5mo PostTransp CD45 BFP-; Mus musculus; RNA-Seq
Freshly CD45 bone marrow cells were isolated by flushing the inner cavity of the long bones from 4.5 months post-transplanted NSG mice. Cell flushing was performed with a typical 1mL syringe containing 1mL DPBS (Gibco). Bone marrow cells were erythrocyte-depleted by incubation with  ACK lysis buffer (Gibco, Cat #A1049201) for 10 minutes at room temperature. Bone marrow cells were stained with anti-mouse CD45-FITC antibody (BD Pharmingen, Cat #553079), and CD45+ BFP+ and CD45+ BFP- cells were isolated by FACS into DPBS suplemented with 0.04% BSA to to a final concentration of 700-1200 cells/μl per sample.  Number of cells encapsulated varied from 10000 to 14000 depending on the sample, to achieve a final target cell recovery of 5000-7000 cells. Library was performed according to the manufacturer's instructions (single cell 3' v3 protocol, 10x Genomics). Resulting libraries were quantified and quality checked by using the 2200 TapeStation® (Agilent).