Team:Calgary/Notebook/Protocols/Process8

From 2011.igem.org

(Difference between revisions)
Line 6: Line 6:
Total DNA isolation protocol
Total DNA isolation protocol
-
<ul>
+
<ol>
<li>Place 1.5 ml of algae culture into centrifuge tube.</li>
<li>Place 1.5 ml of algae culture into centrifuge tube.</li>
<li>Spin down cells at 5000 rpm for 1 minute.</li>
<li>Spin down cells at 5000 rpm for 1 minute.</li>
Line 24: Line 24:
<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>
-
</ul>
+
</ol>
</html>
</html>
}}
}}

Revision as of 22:46, 28 September 2011


Nuclear DNA extraction from Algae

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

Retrieved from "http://2011.igem.org/Team:Calgary/Notebook/Protocols/Process8"