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Simplified Agrobacterium-mediated Transformation of Arabidopsis thaliana
Published: Mar 5, 2011 DOI: 10.21769/BioProtoc.41 Views: 21435
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Abstract
Acquiring transgenic lines is fundamental strategy for plant gene/protein function study. How to successfully obtain the transgenic lines is a big matter with transformation methods. Now Arabidopsis floral dip transformation method is more routinely used in hundreds of laboratories because of easily operating and high successful rate. Here simplified Arabidopsis transformation protocol was described, which was commonly used in our lab and developed from several protocols.
Materials and Reagents
- Arabidopsis plants
- Agrobacterium strain(s): ABI (refs. 16, 17), GV3101 (ref. 16), EHA105 (ref. 18), LBA 4404 (ref. 19) or others
- Murshige&Skoog salts (MS) (Phytotechnology labs.TM)
- Tryptone
- Yeast extract
- NaCl
- Carbenicillin
- Hydrochloric acid
- 5% (wt/vol) sucrose, prepared fresh (Sigma-Aldrich)
- Silwet L-77 (Lehle Seeds)
- Bleach (Thermo Fisher Scientific)
- Appropriate antibiotics (Sigma-Aldrich/ Thermo Fisher Scientific)
- 1/2 MS medium (see Recipes)
- Liquid LB medium (see Recipes)
Equipment
- Growth chambers or light room or greenhouse, adjustable to long-day condition of 16 h light/8 h dark
- Plant pots: e.g., 3.5 in x 3.5 in
- Laminar-flow hood
- Petri dishes (100 x 15 cm VWR)
- Whatman filter paper
- Desiccator jar
Procedure
Category
Molecular Biology > DNA > Transformation
Plant Science > Plant transformation > Agrobacterium
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