American Society of Hematology

Fig. 5.

$Fig. 5. PCR analysis of posttransplantation samples on animal L161. L161 received cells that were transduced (G1Na) and transduced (LNL6) and expanded in the presence of IL-3, IL-6, SCF, and Flt-3 ligand. PB mononuclear cells (PBMNCs), granulocytes (PB GRANS), and BM mononuclear cells (BMMNCs) were analyzed by PCR for the neogene. The 16-bp difference between the vectors in the amplified neoregion allows simultaneous assessment of marking in progeny from the nonexpanded (G1Na) and the expanded (LNL6) infused fractions. Concurrent amplification of β-actin was performed on each sample to quantitate the amount of amplifiable template DNA. Serial dilutions and mixtures of DNA from the producer cell lines containing a known copy number of the neo gene into control rhesus PB DNA at the indicated percentages were used to quantitate the percentage of marked cells posttransplant. RQ1255 represents a sample simultaneously processed and extracted from a negative control animal.$

PCR analysis of posttransplantation samples on animal L161. L161 received cells that were transduced (G1Na) and transduced (LNL6) and expanded in the presence of IL-3, IL-6, SCF, and Flt-3 ligand. PB mononuclear cells (PBMNCs), granulocytes (PB GRANS), and BM mononuclear cells (BMMNCs) were analyzed by PCR for the neogene. The 16-bp difference between the vectors in the amplified neoregion allows simultaneous assessment of marking in progeny from the nonexpanded (G1Na) and the expanded (LNL6) infused fractions. Concurrent amplification of β-actin was performed on each sample to quantitate the amount of amplifiable template DNA. Serial dilutions and mixtures of DNA from the producer cell lines containing a known copy number of the neo gene into control rhesus PB DNA at the indicated percentages were used to quantitate the percentage of marked cells posttransplant. RQ1255 represents a sample simultaneously processed and extracted from a negative control animal.

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