Mapping by Multiple Endonuclease Digestions

Kenneth D. Bloch1

1 Harvard Medical School, Boston, Massachusetts
Publication Name:  Current Protocols in Molecular Biology
Unit Number:  Unit 3.2
DOI:  10.1002/0471142727.mb0302s13
Online Posting Date:  May, 2001
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Abstract

In this unit, a protocol is provided in which the DNA of interest is cleaved with a variety of restriction endonucleases, either individually or in combination, and the resulting products are separated by agarose or acrylamide gel electrophoresis. By determining the sizes of DNA fragments produced by endonuclease cleavage, the restriction map is deduced progressively from simple situations where enzymes cleave the DNA once or twice to more complex situations where cleavage occurs more frequently.

     
 
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Table of Contents

  • Restriction Mapping
  • Commentary
  • Examples
  • Figures
     
 
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Materials

Basic Protocol 1:

  Materials
  • Restriction endonucleases (Table 97.80.4711)
  • 10× restriction endonuclease buffers (unit 3.1)
  • Additional reagents and equipment for agarose or polyacrylamide gel electrophoresis (units 2.5 or 2.7) and DNA molecular weight markers (units 2.5)
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Figures

Videos

Literature Cited

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