GSM8001806: B6_12M_20M_PBMCs_scRNA; Mus musculus; RNA-Seq
Whole blood was collected from 12M and 20M SM/J and BL6 mice via intracardiac puncture and diluted into an equal volume of Dulbecco's PBS + 2% FBS (StemCell Technologies, 07905). According to the manufacturer's specifications, blood was layered on top of Lymphoprep™ (StemCell Technologies, 07851) in a 15 mL tube and centrifuged at 800 rcf for 20 minutes at room temperature. The mononuclear cells were washed twice with PBS + 2% FBS, and platelets were removed with an additional centrifugation at 120 rcf for 10 minutes. To facilitate removal of any remaining RBCs, a lysis step was performed (Biolegend, 420302). Briefly, lysis buffer (1X) was added to the cell suspension and incubated for 5 minutes, with periodic inversion. Cells were then centrifuged at 350 rcf for 10 minutes, and PBMCs were washed twice with PBS + 2% FBS. Libraries were constructed according to manufacturer's specification for the 10X Genomics 3' single-cell gene expression analysis, with multiplexing