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Biol. 53, 159 (1970). 4 S. N. Cohen, A. C. Y. Chang, and L. Hsu, Proc. Natl. Acad. Sci. A. 69, 2110 (1972). 5 R. Benzinger, I. Kleber, and R. Huskey, J. Virol. 7, 646 (1971). e L. Lawhorne, I. Kleber, C. Mitchell, and R. Benzinger, J. Virol. 12, 733 (1973). W. D. Henner, I. Kleber, and R. Benzinger, J. Virol. 12, 741 (1973). 1 ml. 1 ml of spheroplasts are added. 1 ml of fresh E. 4M sucrose, 5 mM MgSO4, and 1 mM CaCl2 in tryptone broth (warmed to 45°) are added, and the mixture is plated in the usual manner.
Possible variations in these conditions are described below. A typical reaction mixture contains 63 mM Tris-HC1 (pH 4 s. s. Price, J. M. Schwing, and P. T. Englund, J. Biol. Chem. 248, 7001 0973). s p. T. Englund, S. S. Price, J. M. Schwing, and P. H. Weigel,in "DNA Synthesis/n Vitro" (R. D. Wells and R. B. ), p. 35. Univ. Park Press, Baltimore, Maryland, 1973. sa j. F. Burd and R. D. Wells, J. Biol. Chem. 249, 7094 0974). 6 p. Modrich and R. A. Rubin, J. Biol. Chem. 252, 7273 (1977). 7 p. H. Weigel, P.
Grece, this series, Vol. 12, Part A, p. 533. 1~D. Smith and H. O. Halvorson, this series, Vol. 12, Part A, p. 538. 13M. Eisenstadt and G. Brawerman, this series, Vol. 12, Part A, p. 541. 14K. Miura, this series, Vol. 12, Part A, p. 543. 15M. G. Smith, this series, Vol. 12, Part A, p. 545. 16H. Stem, this series, Vol. 12, Part B, p. 108. 17K. S. Kirby, this series, Vol. 12, Part B, p. 87. 46 TECHNIQUES FOR LABELING TERMINI [7] solution in 2 volumes of ethanol can be chilled in a Dry Ice-ethanol bath for 15 min and centrifuged in an Eppendoff Microcentrifuge (Brinkman Company) for 10 min.