Two independent LT-HSC clones contribute most of the hematopoiesis seen in mouse L1.

A portion of the bone marrow used for FACS analysis shown in Figure 6was used to inject secondary lethally irradiated mice for day-8 CFU-S. A total of 50 000 nucleated bone marrow cells were used per injection. Macroscopic spleen colonies were individually dissected 8 days post-reconstitution, and then genomic DNA isolated from each colony was used in Southern analysis to determine each proviral integration site. Whole B, T, and myeloid cell samples were obtained by positive magnetic bead selection (Miltenyi Biotech) using single cell suspensions of spleen (B and myeloid cells) and thymus (T cells).