Materials and Reagents

1. Potato plant (Solanum tuberosum cv. Désirée)
   
2. Potato cyst nematode (G. rostochiensis) cysts
   
3. Sodium azide (RICCA Chemical, catalog number: 7144-16 )
   
4. Mercuric chloride (RICCA Chemical, catalog number: 4650-16 )
   
5. Sterile distilled water
   
6. Gentamycin (Fisher Scientific, catalog number: 61398-0010 )
   
7. Nystatin (Sigma-Aldrich, catalog number: N-3503 )
   
8. Agarose (Fisher Scientific, catalog number: BP160-100 )
   
9. Timentin (PhytoTechnology Laboratories^®, catalog number: T869 )
   
10. Micropore tape (Fisher Scientific, catalog number: 19-027-761 )
    
11. Sterilization solution (0.004% mercuric chloride/0.004% sodium azide)
    
12. Hoagland's solution (Sigma-Aldrich, catalog number: H2395 )
    
13. Amberlite XAD-4 resin (Fisher Scientific, catalog number: AC20223 )
    
14. MS salt (Caisson Laboratories, catalog number: MSP01-50LT )
    
15. Inositol (Fisher Scientific, catalog number: AC122261000 )
    
16. Thiamine HCl (Sigma-Aldrich, catalog number: T1270 )
    
17. Gelrite (Fisher Scientific, catalog number: CAS 71010-52-1 )
    
18. Potato root diffusate (PRD) (see Recipes)
    
19. Propagation medium (see Recipes)
    
20. 0.1% agarose (see Recipes)
    

Equipment

1. Sieves No. 60 (250 μm), No. 200 (75 μm) and No. 500 (25 μm) (Endecotts, catalog number: 683927 , 681837 , 691131 )
   
2. Sterile nematode egg hatching chamber (stainless steel metal pan cover and collection pan may be purchased from Vollrath Products, size six inches; the metal screened pan cover with wire-screen in the central area as shown in Figure 3 was custom made)
   
3. Sterile 6-well plates (Greiner Bio-One GmbH, catalog number: 657185 )
   
4. Stirplate
   
5. Sterile forceps and scalpel
   
6. Growth incubator
   
7. 30-μm mesh (Small Parts, catalog number: 7050-1220-000-12 )
   
8. Tissue culture biosafety cabinet
   
9. Centrifuge
   
10. Incubator shaker (New Brunswick Scientific, model: C24KC )
    
11. S. tuberosum growth chambers (we use several types of growth chambers such as I-66LLVL from Percival)
    
12. Rubber stopper (size no. 1) (Fisher Scientific, catalog number: 14-130C )
    
13. Beaker
    
14. Aluminum foil
    
15. Dissecting microscope
    

Procedure