McClean: Pouring Gels for Electrophoresis

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Contents

Overview

The normal method of pouring gels wastes agarose if you only need to run a few lanes. Instead, by preparing gels on double-wide microscope slides, you're able to prolong the life of your agarose stock by quite a while.

Materials

  • Prepared Agarose
  • 75 mm by 50 mm microscope slides (Sigma CLS294775X50 or equivalent)
  • One 10 mL pipet
  • Gel comb with 2" binder clips attached to each end
  • A manual (read: non-electric) pipet filler


Procedure

  1. After heating the agarose, let cool until warm, but not hot, to the touch (similar to when we pour plates)
  2. Meanwhile, adjust the comb so that it is just above the slide. Best way to do this is to layer another slide on top of the first and adjust the comb and binder clips so that the comb is firmly resting on the top slide. Remove this slide and continue.
  3. Adjust the comb over the slide so that it is parallel to and about a centimeter down from the top of the slide. Try to avoid having the teeth of the comb partially hang over the edges of the slide if you can help it.
  4. Fill the pipet with agarose to just over 10 mL and, while maintaining even flow, pour on the slide until about half of a mL remains. Letting the pipet empty fully runs the risk of individual drops of agarose falling on the gel, disrupting surface tension, and making a mess.
  5. When the gel is cool, pull the comb out gently while holding the edges of the gel with your fingers.
  6. Put the gel, still on the slide, in the gel box and load. Make sure it is straight before you turn the power on.

Notes

Please feel free to post comments, questions, or improvements to this protocol. Happy to have your input!

  1. List troubleshooting tips here.
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  3. Anecdotal observations that might be of use to others can also be posted here.


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References

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