GSM9546335: Control Mouse, replicate 2; Mus musculus; RNA-Seq
Mice were humanely euthanized following approved institutional protocols. Bone marrow was harvested by carefully isolating and crushing the femur bones using sterile instruments. The resulting bone marrow suspension was filtered through a 40 µm cell strainer to remove debris and then treated with ammonium chloride potassium (ACK) lysis buffer to lyse and eliminate red blood cells. The remaining bone marrow suspension, free of red blood cells, was washed with phosphate-buffered saline (PBS) and resuspended in a single-cell solution. For single-cell RNA sequencing (scRNA-seq), the prepared cell suspension was processed using the 10x Genomics Chromium Single Cell 3' workflow, following the manufacturer's protocol. Library preparation included reverse transcription, cDNA amplification, and barcoding of individual cells, enabling high-throughput sequencing and downstream analysis of gene expression at the single-cell level.