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Extra resources for Nucleic Acids Part I
65 Copyright © 1980by AcademicPress, Inc. All rights of reproduction in any form reserved. ISBN 0-12-181965-5 28 ASSAYS F O R [CLASS I I R E S T R I C T I O N ] E N D O N U C L E A S E S  breaks with this DNA. These results indicate that the exonuclease is present in excess so that a DNA molecule is essentially digested completely to acid-soluble fragments immediately upon receiving one doublestranded restriction cleavage. The exonuclease-coupled assay described here is well suited to the purification and quantitation of restriction enzymes, since it is simpler to perform and quantitate than an agarose gel electrophoresis assay.
3, 863 (1976). 3 H. K6ssel and R. Roychoudhury, Eur. J. Biochem. 22, 271 (1971). aa R. Roychoudhury and H. K6ssel, Eur. J. Biochem. 22, 310 (1971). 4 R. Roychoudhury, J. Biol. Chem. 247, 3910 (1972). H. K6ssel, R. Roychoudhury, D. Fischer, and A. Otto, Voi. 29, p. 322. o R. Roychoudhury, D. Fischer, and H. K~issel,Biochem. Biophys. Res. Commun. 45, 430 (1971). s D. A. Jackson, R. H. Symons, and P. Berg, Proc. Natl. Acad. Sci. A. 69, 2904 (1972). E E. Lobban and A. D. Kaiser, J. Mol. Biol. 78, 453 (1973).
Mol. Biol. 66, 209 (1972). METHODSIN ENZYMOLOGY,VOL. 65 Copyright © 1980 by Academic Press, Inc. All fightsof reproductionin any form reserved. ISBN 0-12-181965-5 40 TECHNIQUES FOR LABELING TERMINI  1. Radioactivity can be introduced at the 3' terminus of a duplex without altering the structure of the terminus. 4 2. *,5 The extent of r e m o v a l can be precisely predicted if previous sequence data exists. Similarly, one or m o r e nucleotides can be added to a 3' terminus which is base-paired to an appropriate template.