Materials and Reagents

1. Inflorescences from young, healthy Arabidopsis thaliana plants
   
2. Agar plates, with 1/2x Murashige and Skoog solid medium
   
3. Pectolyase (Sigma-Aldrich, catalog number: P3026 )
   
4. Cellulase (Sigma-Aldrich, catalog number: C1184 )
   
5. β-glucoronidase (Roche Diagnostics, catalog number: 03707598001 )
   
6. Sucrose (Affymetrix, catalog number: 21938 )
   
7. Ethanol (EMD Millipore, catalog number: 1085430250 )
   
8. Chloroform (EMD Millipore, catalog number: 102442 )
   
9. Glacial acetic acid (EMD Millipore, catalog number: 100063 )
   
10. Citric acid anhydrous (Affymetrix, catalog number: 13729 )
    
11. Sodium citrate dihydrate (Affymetrix, catalog number: 13735 )
    
12. Formamide (Sigma-Aldrich, catalog number: F9037 )
    
13. Saline-sodium citrate (SSC) (Sigma-Aldrich, catalog number: S0902 )
    
14. Dextran sulfate (Sigma-Aldrich, catalog number: 42867 )
    
15. Salmon sperm (Life Technologies, catalog number: 15632-011 )
    
16. VECTASHIELD^® mounting medium with DAPI (10 ml) (Vector Laboratories, catalog number: H-1400 )
    
17. Fluorescein High-Prime labeling kit (Roche Diagnostics, catalog number: 11585622910 )
    
18. pAL1 centromere probe (ABRC, catalog number: CD3-16 )
    
19. Taq DNA polymerase with standard Taq buffer (New England Biolabs, catalog number: M0273S )
    
20. QIAquick Gel Extraction Kit (kit for purification of DNA from gels) (QIAGEN, catalog number: 28704 )
    
21. Gel Pilot 1 Kb Ladder (molecular weight marker for DNA) (QIAGEN, catalog number: 239085 )
    
22. Agarose for electrophoresis (Bio-Rad Laboratories, catalog number: BR 161-3100 )
    
23. Carnoy’s solution (see Recipes)
    
24. 10 mM Sodium citrate buffer (see Recipes)
    
25. Enzyme digestion buffer (see Recipes)
    
26. FISH buffer (see Recipes)
    
27. 10x PBS (see Recipes)
    
28. 20x SSC (see Recipes)
    

Equipment

1. Hypodermic needles and syringe (Terumo Medical Corporation, model: 26G x ½”, catalog number: NN-2613R )
   
2. Tweezers (Dumont)
   
3. Poly-Prep^® poly-L-lysine coated slides (Sigma-Aldrich, catalog number: P0425-72EA )
   
4. Nunc^® Lab-Tek^® II slides (Thermo Fisher Scientific, catalog number: 154453 )
   
5. Coverslips (Matsunami Glass, catalog number: C218181 )
   
6. Hybridisation oven OV1 (Biometra, catalog number: 052-090 )
   
7. Thermal cycler (Biometra, catalog number: 050-551 )
   
8. Dry block (Thermo Fisher Scientific, Reacti-Therm^®)
   
9. -86 °C refrigerator (Thermo Fisher Scientific, model: Forma^® 88000 )
   
10. Frigidaire -20 °C refrigerator (Frigidaire, model: FCFS201LFB )
    
11. Fluorescence microscope
    
12. Coplin staining jars (Thermo Fisher Scientific, catalog number: 107 )
    
13. Rubbermaid LunchBlox^® food containers
    
14. Super Pap Pen’ liquid blocker pen (Electron Microscopy Sciences, catalog number: 71310 ) (optional)
    

Procedure