Agarose gel electrophoresis

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*If you are getting unexpected bands on your gel you may want to look at the [[Agarose gel electrophoresis/Common issues | common issues in agarose gel electrophoresis page]].
*If you are getting unexpected bands on your gel you may want to look at the [[Agarose gel electrophoresis/Common issues | common issues in agarose gel electrophoresis page]].
*If you have no experience with gel electrophoresis or are explaining it to someone new, [http://arbl.cvmbs.colostate.edu/hbooks/genetics/biotech/gels/virgel.html here] is a cute java demo of what happens.
*If you have no experience with gel electrophoresis or are explaining it to someone new, [http://arbl.cvmbs.colostate.edu/hbooks/genetics/biotech/gels/virgel.html here] is a cute java demo of what happens.
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[[Category:Protocol]]

Revision as of 11:26, 10 July 2006

Contents

General Information

General Procedure

  1. Cast a gel
  2. Place it in gel box in running buffer
  3. Load samples
  4. Run the gel
  5. Image the gel

Buffers

  • TAE - better resolution of fragments >4kb;
  • TBE - better resolution of 0.1-3kb fragments; TBE is better suited for high-voltage (>150V) electrophoresis than TAE because of its higher buffering capacity and lower conductivity;

Loading dyes

dye 0.5-1.5% agarose 2.0-3.0% agarose*
xylene cyanol 4000-5000 bp 750 bp
bromophenol blue 400-500 bp 100 bp

*sieving agarose

Specific Protocols

Endy:Agarose gel

Knight:Agarose gel electrophoresis

Notes

Personal tools