Viral particles were generated by transient co-transfection of the Phoenix-Eco packaging cell line with the retroviral construct and the pCL-Eco plasmid (Imgenex, San Diego, CA). Viral supernatants were harvested at two and three days after transfection, and immediately frozen at -80°C until use. For experiments involving retroviral transduction (Figs. 6 –7 ), tissue culture plates (Costar, Corning) were incubated overnight with 33 µg/ml Retronectin (Clontech) and 2 µg/ml of DL1-ext-IgG protein, and then loaded with viral supernatant. Cells were then cultured directly on virus-bound plates under the conditions described below. For experiments involving culture of sorted progenitors on DL1-coated plates (Fig. 6 ), tissue culture plates (Costar, Corning) were incubated overnight with 33 µg/ml Retronectin (Clontech) with different concentrations of the DL1-ext-IgG protein, as indicated.
Pcl eco plasmid
Manufactured by Novus Biologicals
The PCL-Eco plasmid is a vector designed for use in molecular biology research. It contains an origin of replication and antibiotic resistance genes, allowing for selection and propagation in bacterial host cells.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using pcl eco plasmid
1
Retroviral Transduction of Cells
2
Retroviral Transduction of Cells
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Viral particles were generated by transient co-transfection of the Phoenix-Eco packaging cell line with the retroviral construct and the pCL-Eco plasmid (Imgenex, San Diego, CA). Viral supernatants were harvested at two and three days after transfection, and immediately frozen at -80°C until use. For experiments involving retroviral transduction (Figs. 6 –7 ), tissue culture plates (Costar, Corning) were incubated overnight with 33 µg/ml Retronectin (Clontech) and 2 µg/ml of DL1-ext-IgG protein, and then loaded with viral supernatant. Cells were then cultured directly on virus-bound plates under the conditions described below. For experiments involving culture of sorted progenitors on DL1-coated plates (Fig. 6 ), tissue culture plates (Costar, Corning) were incubated overnight with 33 µg/ml Retronectin (Clontech) with different concentrations of the DL1-ext-IgG protein, as indicated.
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