Materials and Reagents

1. Recipient strain [ATCC2001, HTL or clinical isolates (Schwarzmuller et al., 2014)]
   
2. DNA deletion construct / transforming DNA (highly purified)
   
3. Electroporation cuvette (Life Technologies, Invitrogen^TM, catalog number: P51050 )
   This product has been discontinued.
   
4. Deep well plate (96-well) (Nunc^®, catalog number: 7322662 )
   
5. Reservoir (autoclavable) (VWR International, catalog number: 6130466 )
   
6. Multichannel pipette (200 µl) (BrandTech Scientific)
   
7. Sterile water (double-distilled)
   
8. Bacto™ peptone (BD Biosciences, catalog number: 211820 )
   
9. Bacto™ yeast extract (BD Biosciences, catalog number: 212720 )
   
10. Bacto™ agar (BD Biosciences, catalog number: 214030 )
    
11. Glucose (Merck KGaA, catalog number: 108337)
    
12. Tris base (Sigma-Aldrich, catalog number: T1503 )
    
13. EDTA (Sigma-Aldrich, catalog number: E6758 )
    
14. Lithium acetate dihydrate (Sigma-Aldrich, catalog number: L6883 )
    
15. DL-Dithiothreitol (DTT) (Sigma-Aldrich, catalog number: D0632 )
    
16. Nourseothricin-dihydrogen sulfate (Werner BioAgents, catalog number: 5.0 )
    
17. TE buffer (see Recipes)
    
18. DTT solution (see Recipes)
    
19. YPD media (see Recipes)
    
20. Solid selective media (see Recipes)
    

Equipment

1. 96-wel electroporation plate (2 mm gap) (Harvard Apparatus, catalog number: HTP962450450 )
   
2. Electroporation cuvettes (1 mm gap) (Life Technologies, Invitrogen^TM, catalog number: P51050 )
   This product (catalog P51050 ) has been discontinued.
   
3. Culture flasks with baffles
   
4. Centrifuge for tubes (50 ml) (Eppendorf, catalog number: 5702R )
   
5. Centrifuge for tubes (1.5 ml) (Eppendorf, catalog number: 5417R )
   
6. Rotary shaker for culture flasks (New Brunswick Scientific, catalog number: innova44 )
   
7. Stratagene Stratalinker 2400 UV Crosslinker (Artisan Technology Group^®, catalog number: 532741 )
   
8. Single-well scaleTransfection apparatus consisting of capacitance extender, pulse controller and gene pulser (Bio-Rad Laboratories, AbD Serotec^®, catalog numbers: 165-2087 ; 165-2098 ; 165-2077 )
   
9. 96-well microtiter plates
   
   1. 96-well electroporation device (BTX The Electroporation Experts, catalog number: ECM63 or Harvard Apparatus, catalog number: 450422 )
      
   2. Plate handler (BTX The Electroporation Experts, catalog number: HT-100 or Harvard Apparatus, catalog number: 450400 )

Procedure