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Culture 293T cells at 1.5 – 2.0 X106 cells/ 4 mL of DIO in a 60mm petri dish. Day 2 Calcium phosphate transfection of 293T cells (cells should be approximately 80 percent confluent) In a 5mL sterile tube, combine 400 (l ddH2O, 100 (l 1.25 M CaCl2. Add DNA. Retroviral DNA 7-10(g . PCLEco 3-5 (g *maximum of 12(g total ................
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