1. Harvest of supernatant containing retrovirus:
a. 0.45 |im Disposable sterile syringe filters with cellulose acetate membrane.
b. Cryogenic vials with gasket seal closure.
2. Enzymatic stain for lacZ activity:
a. 0.1 M sodium phosphate buffer, pH 7.2.
b. Fixative: 4% paraformaldehyde, 0.5% glutaraldehyde in 0.1 M sodium phosphate buffer, pH 7.2. For 400 mL: Heat 200 mL of 0.1 M sodium phosphate buffer and 16 g of paraformaldehyde in a fume hood with stirring until dis solved (see Note 3). Add 8 mL of 25% glutaraldehyde stock and the remaining 192 mL of 0.1 M sodium phosphate buffer. Store at 4°C.
c. 40 mg/mL 5-bromo-4-chloro-3-indolyl-P-d-galactopyranoside (X-gal) in DMSO. Store at -20°C.
d. Staining buffer (see Note 4): 5 mM K3Fe(CN)6. 5 mM K4Fe(CN)63H2O; 2 mM MgCl2 in PBS, pH 7.2-7.4 (see Note 5). Store at 4°C.
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