Digestion of DNA with Restriction Endonucleases

Kenneth D. Bloch1

1 Massachusetts General Hospital, Charleston, Massachusetts
Publication Name:  Current Protocols in Neuroscience
Unit Number:  Appendix 1M
DOI:  10.1002/0471142301.nsa01ms11
Online Posting Date:  May, 2001
GO TO THE FULL TEXT: PDF or HTML at Wiley Online Library

Abstract

Restriction endonucleases recognize short DNA sequences and cleave double‐stranded DNA at specific sites within or adjacent to the recognition sequences. Restriction endonuclease cleavage of DNA into discrete fragments is one of the most basic procedures in molecular biology. This appendix describes restriction endonucleases and their properties.

     
 
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library

Table of Contents

  • Basic Protocol 1: Digesting a Single DNA Sample with a Single Restriction Endonuclease
  • Reagents and Solutions
  • Literature Cited
  • Tables
     
 
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library

Materials

Basic Protocol 1: Digesting a Single DNA Sample with a Single Restriction Endonuclease

  Materials
  • DNA sample in H 2O or TE buffer
  • Restriction endonuclease buffers (see reciperecipes)
  • Restriction endonucleases
  • Loading buffer, 10×: 20% (w/v) Ficoll 400/0.1 M Na 2 EDTA (pH 8.0)/1.0% (w/v) SDS/0.25% (w/v) bromphenol blue/0.25% (w/v) xylene cyanol (optional)
  • 0.5 M EDTA, pH 8.0 ( appendix 2A; optional)
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library

Figures

Videos

Literature Cited

Literature Cited
   Danna, A.J. 1980. Determination of fragment order through partial digests and multiple enzyme digests. Methods Enzymol. 65:449‐467.
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library