Agarose gel electrophoresis

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Agarose gel electrophoresis is used to separate DNA or RNA molecules by size. Nucleic acids are negatively charged and are moved through an agarose matrix by an electric field (electrophoresis). Shorter molecules move faster and migrate further.

Contents

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

for recipes see: Agarose gel loading dye

Notes

See also

Specific Protocols

External links

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