Mapping by Partial Endonuclease Digestions

Kenneth D. Bloch1

1 Harvard Medical School, Boston, Massachusetts
Publication Name:  Current Protocols in Molecular Biology
Unit Number:  Unit 3.3
DOI:  10.1002/0471142727.mb0303s09
Online Posting Date:  May, 2001
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Abstract

In the procedure described in this unit, a DNA fragment radiolabeled at one of its two ends is purified by gel electrophoresis and subjected to partial cleavage by a restriction endonuclease. Analysis of the resulting products by polyacrylamide (or agarose) gel electrophoresis enables one to define the distance of restriction sites from the labeled end.

     
 
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Table of Contents

  • Commentary
  • Examples
  • Key References
     
 
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Materials

Basic Protocol 1:

  Materials
  • Reagents and equipment for digesting DNA with restriction enzymes (unit 3.1), labeling DNA (unit 3.5 or ), agarose or polyacrylamide gel electrophoresis (unit 2.5 or ), and autoradiography ( appendix 3A)
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Figures

Videos

Literature Cited

Key References
   Boseley, P.G., Moss, T., and Birnstiel, M.L. 1980. 5′ labeling and poly(dA) tailing. Meth. Enzymol. 65:478‐494.
   Danna, A.J. 1980. Determination of fragment order through partial digests and multiple enzyme digests. Meth. Enzymol. 65:449‐467.
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