Materials and Reagents

1. R. capsulatus recipient strain
   
2. Escherichia coli donor strain (containing plasmid to be conjugated) capable of conjugation (e.g. S17-1) or E. coli donor strain containing plasmid to be conjugated and a helper strain containing the tra genes [e.g. HB101 (pRK2013)]. For a review on conjugation and tra genes see Willetts et al. (1984)
   
3. Plasmid to be conjugated into R. capsulatus (e.g. pXCA601; Tetracycline resistance)
   
4. Appropriate antibiotic (resistance specified by plasmids and bacterial strains)
   
5. Thiamine hydrochloride
   
6. H₃BO₃
   
7. MnSO₄·H₂O
   
8. Na₂MoO₄·2H₂O
   
9. ZnSO₄·7H₂O
   
10. Cu(NO₃)·3H₂O
    
11. D, L-malic acid
    
12. Na₂EDTA
    
13. MgSO₄·7H₂O
    
14. CaCl₂·2H₂O
    
15. FeSO₄·7H₂O
    
16. 10 mM potassium phosphate buffer
    
17. 0.3% Difco yeast extract
    
18. 0.3% Difco bactopeptone
    
19. Bacto-tryptone
    
20. Yeast extract
    
21. Trace element solution (see Recipes)
    
22. RCV broth (see Recipes)
    
23. RCV agar (see Recipes)
    
24. LB broth (see Recipes)
    
25. LB agar (see Recipes)
    
26. YPS agar (see Recipes)

Equipment

1. 30 °C and 37 °C shakers
   
2. 30 °C and 37 °C incubator
   
3. Test tubes
   
4. Petri plates
   
5. Sterile 1.7 ml microcentrifuge tubes
   
6. Inoculation loop
   
7. Pipetmen (10 μl to 1 ml range) and appropriate tips
   
8. Graduated pipette (5 ml range) and aspiration bulb
   
9. Bench-top microcentrifuge with rotor for 1.7 ml microcentrifuge tubes
   

Procedure