Methods in virology / Vol.4 by Karl Maramorosch; Hilary Koprowski

By Karl Maramorosch; Hilary Koprowski

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1967). Proc. Nat. Acad. Sci. U. S. 58, 127. Littlefield, J. W . (1964). Science 145, 709. , a n d Littlefield, J. W. (1968). J. Virol. 2, 69. M a r s t o n , R. Q. (1958). Proc. Soc. Exp. Biol. Med. 98, 853. , a n d Basilico, C. (1968). Nature (London) 220, 1199. Migeon, B. , a n d Miller, C. S. (1968). Science 162, 1005. Milgrom, F . , B a r r o n , A. , a n d Witebsky, E. (1964). J. Immunol. 92, 8. , a n d Bodmer, W. (1969). Nature (London) 223, 358. Neff, J. , a n d Enders, J. F . (1968). Proc.

Its lethal effect is probably largely due to its incorpora­ tion into D N A . Cells grown in medium containing the analog at doses of a b o u t 10 jug/ml do not die rapidly; they enlarge, division ceases, and they may take a week or more to detach from the glass or plastic. Cells lacking thymidine kinase ( T K " cells) are completely resistant to B U d R . They have been obtained by treatment over several months with gradually increasing doses of B U d R (see, for example, Marin and Littlefield, 1968).

The gradient may be formed well in advance and the material to be fractionated needs only to be inserted in the cavity im­ mediately prior to the acceleration of the rotor. This nylon reograd rotor is admirably suited to the crude fractionation 48 A. POLSON of a mixture of macromolecular substances, but if small fractions of high purity are required, they may be obtained by isopycnic centrifugation in the multi-partition cell of the analytical ultracentrifuge. A n account of this follows. E . M U L T I - P A R T I T I O N C E L L FOR ISOPYCNIC SEPARATION IN D E N S I T Y G R A D I E N T S IN THE A N A L Y T I C A L U L T R A C E N T R I F U G E Katz et al.

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