Team:Calgary/Notebook/Protocols/Process8
From 2011.igem.org
Revision as of 04:14, 29 September 2011 by Rpgguardian (Talk | contribs)
![](https://static.igem.org/mediawiki/2011/c/c1/UCalgary_CircleHome.png)
![](https://static.igem.org/mediawiki/2011/d/d4/UCalgary_CircleTeam.png)
![](https://static.igem.org/mediawiki/2011/7/7d/UCalgary_CircleProject.png)
![](https://static.igem.org/mediawiki/2011/8/8d/UCalgary_CircleParts.png)
![](https://static.igem.org/mediawiki/2011/0/03/UCalgary_CircleNotebook.png)
![](https://static.igem.org/mediawiki/2011/8/84/UCalgary_CircleOutreach.png)
![](https://static.igem.org/mediawiki/2011/3/3c/UCalgary_CircleSponsors.png)
![](https://static.igem.org/mediawiki/2011/8/85/UCalgary_CircleiGEM.png)
![](https://static.igem.org/mediawiki/2011/4/42/UCalgary_TitleBar.png)
Nuclear DNA Extraction from Algae
![](https://static.igem.org/mediawiki/2011/e/ed/UCalgary_BodyBoxTop.png)
Total DNA Isolation Protocol
Reagents and Materials
Extraction buffer (for 40 ml total):
1 M Tris HCl pH 7.5 | 8 ml |
5 M NaCl | 2 ml |
0.5 M EDTA | 2 ml |
20% SDS | 1 ml |
ddH2O | 27 ml |
Elution buffer (provided in kit from Qiagen)
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.
![](https://static.igem.org/mediawiki/2011/c/cb/UCalgary_BodyBoxBottom.png)