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This is a simple protocol for isolating genomic DNA from fresh plant tissues. DNA from this experiment can be used for all kinds of genetics studies, including genotyping and mapping. This protocol uses Edward’s extraction buffer to isolate DNA.

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[Bio101] Extract Genomic DNA from Arabidopsis Leaves (Can be Used for Other Tissues as Well)

Molecular Biology > DNA > DNA extraction
Author: Yongxian Lu
Yongxian LuAffiliation: Carnegie Institution for Science, Stanford University, Stanford, USA
For correspondence: yxlu@stanford.edu
Bio-protocol author page: a28
7/5/2011, 13510 views, 1 Q&A, How to cite
DOI: http://dx.doi.org/10.21769/BioProtoc.90

[Abstract] This is a simple protocol for isolating genomic DNA from fresh plant tissues. DNA from this experiment can be used for all kinds of genetics studies, including genotyping and mapping. This protocol uses Edward’s extraction buffer to isolate DNA.

Materials and Reagents

  1. Ethanol (EtOH)
  2. Isopropanol
  3. NaCl
  4. EDTA
  5. SDS
  6. Tris (pH 7.5)
  7. Edward buffer (see Recipes)

Equipment

  1. Ceramic mortar and pestles
  2. Centrifuges (Eppendorf)
  3. Vortex (VWR International)

Procedure

  1. Add 400 μl Edward extraction buffer to a 1.5 μl tube.
  2. Take 2-3 small leaves or 1 big leaf, grind with pestle (2-3 weeks old).
  3. Vortex 5 sec, set at room temperature until all preps are ready.
  4. Spin at 16,000 rpm for 2 min.
  5. Transfer 300 μl of suspension to a fresh tube.
  6. Add 300 μl of isopropanol at room temperature for 2 min.
  7. Spin 5 min, wash pellet with 70% EtOH, and dry at room temperature.
  8. Resuspend in 100 μl H2O and store at -20 °C.
  9. Use 2 - 4 μl for PCR reaction to validate the presence of target DNA.

Recipes

  1. Edward buffer
    200 mM Tris (pH 7.5)
    250 mM NaCl
    25 mM EDTA
    0.5% SDS

References

  1. Lu, Y., Chanroj, S., Zulkifli, L., Johnson, M. A., Uozumi, N., Cheung, A. and Sze, H. (2011). Pollen tubes lacking a pair of K+ transporters fail to target ovules in Arabidopsis. Plant Cell 23(1): 81-93.


How to cite this protocol: Lu, Y. (2011). Extract Genomic DNA from Arabidopsis Leaves (Can be Used for Other Tissues as Well). Bio-protocol Bio101: e90. DOI: 10.21769/BioProtoc.90; Full Text



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8/12/2014 6:34:03 PM  

Marisa Rosa
UC Berkeley

Should it not be ul instead of ml throughout the procedures?

8/12/2014 11:37:05 PM  

Yongxian Lu (Author)
Carnegie Institution for Science,Stanford University

Yes, you are right. It should be ul, instead of ml. Guess it's a typo by the editorial team. sorry for that.

10/10/2016 11:48:23 AM  

Jianxin Zhao
University of Florida

Can this method could be used for extraction of Medicago or red clover ?

Reply

Please login to post your questions/comments. Your questions will be directed to the authors of the protocol. The authors will be requested to answer your questions at their earliest convenience. Once your questions are answered, you will be informed using the email address that you register with bio-protocol.
You are highly recommended to post your data (images or even videos) for the troubleshooting. For uploading videos, you may need a Google account because Bio-protocol uses YouTube to host videos.

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