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| Untersuchte Arbeit: Seite: 32, Zeilen: 1-11 |
Quelle: Fontanari Krause 2006 Seite(n): 90, 91, Zeilen: 90: last lines - 91: 1-7 |
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| 3.3 Retrovirus Production.
Retrovirus was produced with the constructs mentioned before by individually co-transfecting 15 μg of the construct DNA with equal amounts of Ecopac DNA into 293T cells by using the calcium chloride precipitation method of transfection (Pineault et. al., 2003b). Virus conditioned medium (VCM) was collected 48h after transfection at time intervals of eight hours for forty-eight hours. Production of high-titer helper-free retrovirus was achieved by transducing the ecotropic packaging cell line GP+E86 (Pineault et al., 2003b) with virus conditioned medium (VCM) from 293T transfected cells. Successfully transduced GP+E86 cells were sorted out by FACS using GFP or YFP as a marker 4 days after transduction using standard procedures (Pineault et. al., 2003a). Pineault, N., Buske, C., Feuring-Buske, M., Abramovich, C., Rosten, P., Hogge, D. E., Aplan, P. D., and Humphries, R. K. (2003). Induction of acute myeloid leukemia in mice by the human leukemia- specific fusion gene NUP98-HOXD13 in concert with Meis1. Blood 101, 4529-4538. |
[page 90]
5.10.2. Retrovirus Production Retrovirus was produced with the constructs mentioned above by co-transfecting 30 μg of the construct plasmid DNA with equal amounts of Ecopac DNA into 293T cells using the [page 91] calcium phosphate precipitation method (Pineault et al., 2003). Virus conditioned medium (VCM) was collected 48h after transfection at time intervals of eight hours for forty-eight hours. Production of high-titer helper-free retrovirus was achieved by transducing the ecotropic packaging cell line GP+E86 (Pineault et al., 2003) with virus conditioned medium (VCM) from 293T transfected cells. Successfully transduced GP+E86 cells were sorted out by FACS using GFP as a marker 4 days after transduction using standard procedures (Pineault et al., 2003). Pineault N., Buske C., Feuring-Buske M., Abramovich C., Rosten P., Hogge D. E., Aplan P. D., Humphries R. K. (2003). Induction of acute myeloid leukemia in mice by the human leukemiaspecific fusion gene NUP98-HOXD13 in concert with Meis1. Blood. 101 (11), 4529-38. |
The source is not given. Note that there is only one source "Pineault et al. (2003) listed in the bibliography. Note that in Pineault et al. (2003) (the paper listed in the bibliography) the here documented parallel text cannot be found. |
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