Protocol for reticulocyte enrichment from low-volume human blood samples from alpha-thalassemic and healthy participants

Reticulocyte isolation from peripheral blood is crucial for hematological research. Here, we present a protocol for high-quality reticulocyte enrichment from small blood quantities obtained from alpha-thalassemic and healthy participants. We describe steps for Ficoll and Percoll gradient centrifugation to obtain a reticulocyte-enriched fraction, followed by negative immunomagnetic separation to remove granulocytes and platelets. This technique allows enriched reticulocyte isolation for multi-omics hematological analysis. Additionally, we detail procedures to recover peripheral blood mononuclear cells (PBMCs) and erythrocytes from the original blood sample.