the procedure entails ( i ) the preparation of a crude cell lysate by dounce homogenization of cells 3-5 h post-infection , ( ii ) differential centrifugation to give a 15 000 g 'mitochondrial ' pellet , ( iii ) equilibrium centrifugation on discontinuous sucrose gradients ( friedman et al . 1972 ) to give a membranous band of density 1-16 g/ml , ( iv ) solubilization with triton n-101 and velocity centrifugation to give a 25s solubilized polymerase complex and ( v ) affinity chromatography through an oligo ( dt ) -cellulose matrix bearing immobilized 42s virus particle rna .
C0007634
cells
C0026237
mitochondrial