Rhodobacter capsulatus Gene Transfer Agent Transduction Assay

Materials and Reagents

1. RcGTA donor strain (e.g. R. capsulatus Y262 or Rif^R strain)
   
2. Recipient strain (e.g. R. capsulatus DW5 [photosynthesis mutant] or Rif^S strain to test the transfer of the Rif^R allele)
   
3. Potassium phosphate buffer
   
4. L-malic acid (Sigma-Aldrich, catalog number: 240176 )
   
5. Thiamine hydrochloride (Sigma-Aldrich, catalog number: T1270 )
6. G-buffer (filter sterilize) (see Recipes)
   
7. RCV broth/agar¹ (see Recipes)
   
8. YPS broth/agar³ (autoclaved) (see Recipes)
   

Equipment

1. Variable temperature shaker for test tubes (30-35 °C)
   
2. Anaerobic jars for photosynthetic growth (if using photosynthesis mutant for recipient)
   
3. Light box for photosynthetic growth
   
4. 30 °C incubator
   
5. Test tubes
   
6. 17 ml glass screwcap tubes
   
7. 10 ml plastic culture tubes
   
8. Glass beads
   
9. 0.45 μm pore-size filter
   
10. 10 ml syringe
    

Procedure