Materials and Reagents

1. Surgical tape (3M, catalog number: 15300 )
   
2. Sterile petri dishes
   
3. Seeds: Arabidopsis thaliana (Col-0 ecotype) and tobacco (Nicotiana tabacum cv SR1)
   
4. Bacterial strain: Agrobacterium tumefaciens EHA105
   
5. Sodium hypochlorite (Sigma-Aldrich, catalog number: L099100 )
   Note: The Pricing and availability are not currently available.
   
6. Kanamycin sulfate Streptomyces kanamyceticus (Sigma-Aldrich, catalog number: K0879 )
   
7. Timentin (Yeasen, catalog number: 60230ES07 )
   
8. Agar (Sangon Biotech, catalog number: A100637 )
   
9. Methylester sulfonate (Sangon Biotech, catalog number: MB0341 )
   
10. Sucrose (Sigma-Aldrich, catalog number: E001888 )
    
11. Glucose (Sigma-Aldrich, catalog number: G8270 )
    
12. Ethyl alcohol, Pure (Sigma-Aldrich, catalog number: 459844 )
    
13. Deionized water
    
14. Soil
    
15. Murashige and Skoog (MS) basal medium with vitamin powder (PhytoTechnology, Laboratories^®, catalog number: 15B0519117B ) (1 L) (see Recipes)
    
16. 1/2 MS medium (1 L) (see Recipes)
    
17. MS1 medium (1 L) (see Recipes)
    
18. MS2 medium (1 L) (see Recipes)
    
19. MS3 medium (1 L) (see Recipes)
    
20. Callus-inducing medium (1 L) (see Recipes)
    
21. Shoot-inducing medium (1 L) (see Recipes)
    
22. 22 mM 2, 4-dichlorophenoxy (2, 4-D) stock solution (Sigma-Aldrich, catalog number: 31518 ) (see Recipes)
    
23. 9 mM 3-indoleacetic acid (IAA) (Sigma-Aldrich, catalog number: I2886 ) stock solution (see Recipes)
    
24. 20 mM Kinetin (Sigma-Aldrich, catalog number: K0753 ) stock solution (see Recipes)
    
25. 50 mM 6-(γ, γ-Dimethylallylamino)purine (2-IP) (Sigma-Aldrich, catalog number: D5912 ) stock solution (see Recipes)
    
26. 2 mg/ml 6-benzylaminopurine (6-BA) (Sigma-Aldrich, catalog number: B3408 ) stock solution (see Recipes)
    
27. 1 M Potassium hydroxide (KOH) (Sigma-Aldrich, catalog number: P5958 ) (see Recipes)
    
28. 15% bleach (see Recipes)
    
29. 10% bleach (see Recipes)
    
30. Infection buffer (see Recipes)
    

Equipment

1. Clean bench (horizontal laminar flow)
   
2. Autoclave (ZEALWAY)
   
3. Incubator (25 °C/80 UML) (Percival-scientific, model: CU-41L4 )
   
4. Growth chamber
   
5. Glass culture bottles (130 mm x 100 mm x 62 mm)
   
6. Sterile 5 mm-hole punch
   
7. Scalpel (10 cm in length)
   
8. Forceps (20 cm in length)
   
9. Scissor (15 cm in length)
   

Procedure