Team:DTU-Denmark/Gel preparation and gel electrophoresis

From 2011.igem.org

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== Gel preparation (1% gel) (100 ml of the buffer) ==
== Gel preparation (1% gel) (100 ml of the buffer) ==
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<ol style="list-style-type:lower-roman">
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<ol style="list-style-type:lettr">
<li value="a">100 ml of 1x TBE buffer.</li>
<li value="a">100 ml of 1x TBE buffer.</li>
<li value="b">10 μl of ethidium bomide (10 mg/ml).</li>
<li value="b">10 μl of ethidium bomide (10 mg/ml).</li>
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# Place gel container in the electrophoresis machine, remove combs and cover with the TBE buffer.  
# Place gel container in the electrophoresis machine, remove combs and cover with the TBE buffer.  
Gel electrophoresis:
Gel electrophoresis:
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<ol>
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<ol style="list-style-type:higher-roman">
<li value="i">2μl of DNA sample.</li>
<li value="i">2μl of DNA sample.</li>
<li value="ii">3 μl of distilled water.</li>
<li value="ii">3 μl of distilled water.</li>

Revision as of 14:12, 18 September 2011

Gel preparation (1% gel) (100 ml of the buffer)

  1. 100 ml of 1x TBE buffer.
  2. 10 μl of ethidium bomide (10 mg/ml).
  3. 1 g of agarose.
  4. Assembled gel container.
  1. Mix buffer with agarose and heat in microwave until the solution is clear
  2. Add 10μl of ethidium bromide (10mg/ml).
  3. Pour solution to the gel container and leave it to solidify (30-45 min).
  4. Place gel container in the electrophoresis machine, remove combs and cover with the TBE buffer.

Gel electrophoresis:

  1. 2μl of DNA sample.
  2. 3 μl of distilled water.
  3. 1μl of 6x loading dye.
  4. 4.2 μl of Gene Ruler DNA ladder mix from Fermentas.

It is recommended to use 7V for each cm of the gel length and to run the gel for 45 min.

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