Larry A. McReynolds1, Nicole M. Nichols1

1 New England Biolabs, Ipswich, Massachusetts
Publication Name:  Current Protocols in Molecular Biology
Unit Number:  Unit 3.11
DOI:  10.1002/0471142727.mb0311s96
Online Posting Date:  October, 2011
GO TO THE FULL TEXT: PDF or HTML at Wiley Online Library


Reaction conditions for numerous exonucleases are detailed in this unit along with discussions of potential applications and preferred substrates. Single‐stranded and double‐stranded 5′ → 3′ and 3′ → 5′ exonucleases are included. Curr. Protoc. Mol. Biol. 96:3.11.1‐3.11.7. © 2011 by John Wiley & Sons, Inc.

Keywords: DNases; ssDNA exonucleases; dsDNA exonucleases

PDF or HTML at Wiley Online Library

Table of Contents

  • Introduction
  • Single‐Stranded 3′ → 5′ Exonucleases
  • Single‐Stranded 5′ → 3′ and 3′ → 5′ Exonucleases
  • Double‐Stranded 5′ → 3′ Exonucleases
  • Double‐Stranded 3′ → 5′ Exonuclease
  • Single‐ and Double‐Stranded 5′ → 3′ and 3′ → 5′ Exonuclease
  • Literature Cited
  • Figures
  • Tables
PDF or HTML at Wiley Online Library


PDF or HTML at Wiley Online Library



Literature Cited

Literature Cited
   Berk, A.J. and Sharp, P.A. 1977. Sizing and mapping of early adenovirus mRNAs by gel electrophoresis of S1 endonuclease digested hybrids. Cell 12:721‐732.
   Demple, B., Johnson, A., and Fung, D. 1986. Exonuclease III and endonuclease IV remove 3′ blocks from DNA synthesis primers in H2 O2‐damaged Escherichia coli. Proc. Natl. Acad. Sci. U.S.A. 83:7731‐7735.
   Eichler, D.C. and Lehman, I.R. 1977. On the role of ATP in phosphodiester bond hydrolysis catalyzed by the recBC deoxyribonuclease of Esherichia coli. J. Biol. Chem. 252:499‐503.
   Goff, S. and Berg, P. 1978. Excision of DNA segments introduced into cloning vectors by the poly(dA‐dT) joining method. Proc. Natl. Acad. Sci. U.S.A. 75:1763‐1767.
   Henikoff, S. 1984. Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. Gene 28:351‐359.
   Karu, A., MacKay, V., Goldmark, P., and Linn, S. 1973. The recBC deoxyribonuclease of Escherichia coli K‐12. Substrate specificity and reaction intermediates. J. Biol. Chem. 248:4874‐4884.
   Kerr, C. and Sadowski, P.D. 1972. Gene 6 exonuclease of bacteriophage T7. I. Purification and properties of the enzyme. J. Biol. Chem. 247:305‐310.
   Krishnakumar, S., Zheng, J., Wilhelmy, J., Faham, M., Mindrinos, M., and Davis, R. 2008. A comprehensive assay for targeted multiplex amplification of human DNA sequences. Proc. Natl. Acad Sci. U.S.A. 105:9296‐9301.
   Lehman, I.R. and Nussbaum, A.L. 1964. The deoxyribonucleases of Escherichia coli V. on the specificity of exonuclease I (phosphodiesterase). J. Biol. Chem. 239:26‐28.
   Li, H., Cui, X. and Arnheim, N. 1991. Eliminating primers from completed polymerase chain reactions with exonuclease VII. Nucleic Acids Res. 19:3139‐3141.
   Little, J.W. 1981. Lambda exonuclease. In Gene Amplification and Analysis, Vol. 2: Structural analysis of nucleic acids ( J.G. Chirikjian and T.S. Papas, eds.) pp. 135‐145. Elsevier/North Holland, NY, Amsterdam, Oxford.
   Rogers, S.G. and Weiss, B. 1980. Exonuclease III of Escherichia coli K‐12, an AP exonuclease. Methods Enzymol. 65:201‐211.
   Sanger, F., Nicklen, S., and Coulson, A.R. 1977. DNA sequencing with chain‐terminating inhibitors. Proc. Natl. Acad. Sci. U.S.A. 74:5463‐5467.
   Thomas, K.R. and Olivera, B.M. 1978. Processivity of DNA exonucleases. J. Biol. Chem. 253:424‐429.
   Vales, L.D., Rabin, B.A., and Chase, J.W. 1982. Subunit structure of Escherichia coli exonuclease VII. J. Biol. Chem. 257:8799‐8805.
PDF or HTML at Wiley Online Library