Team:Calgary/Notebook/Protocols/Process8

From 2011.igem.org

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<li>Centrifuge at 13000 rpm for 2 minutes.</li>
<li>Centrifuge at 13000 rpm for 2 minutes.</li>
<li>Discard the supernatant, DNA is in the pellet.</li>
<li>Discard the supernatant, DNA is in the pellet.</li>
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Revision as of 22:45, 28 September 2011


Nuclear DNA extraction from Algae

Total DNA isolation protocol
  • Place 1.5 ml of algae culture into centrifuge tube.
  • Spin down cells at 5000 rpm for 1 minute.
  • Pipette off supernatant.
  • Add 400 µl extraction buffer.
  • Incubate for 15 minutes at 50ºC, inverting tubes several times during incubation.
  • Centrifuge at 13000 rpm for 5 minutes.
  • Transfer supernatant to new tube.
  • Add 400 µl isopropanol.
  • Invert several times.
  • Place on ice for 5 minutes.
  • Centrifuge at 13000 rpm for 10 minutes.
  • Pour off supernatant and wash pellet with 500 µL of 70% ethanol, ensuring to resuspend the pellet.
  • Centrifuge at 13000 rpm for 1 minutes.
  • Decant ethanol and place tubes upside down on paper towel to dry off excess ethanol.
  • Dissolve pellet in 50 µl elution buffer, place tube on bench for 4 hours.
  • Centrifuge at 13000 rpm for 2 minutes.
  • Discard the supernatant, DNA is in the pellet.

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