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Table 1 Consensus clostridial electrotransformation conditions leading to initial low-level transformation of C. pasteurianum

From: Development of an electrotransformation protocol for genetic manipulation of Clostridium pasteurianum

Electrotransformation parameter Consensus for Clostridium species Low-level transformation of C. pasteurianum Selected references
Cell growth    
 Growth medium YTG or 2×YTG 2×YTG [44, 58, 59]
 Growth phase and OD600 at time of harvest mid to late exponential phase (OD600 0.5–0.9) OD600 0.6-0.8 [25]
Washing and pulse delivery    
 Wash and electroporation buffer 5–7 mM sodium phosphate, pH 6.5-7.4, containing 270 mM sucrose and 1 mM MgCl2 5 mM sodium phosphate, pH 6.5, containing 270 mM sucrose and 1 mM MgCl2 [25, 26, 29]
 Number of wash steps 1 1 [44, 45, 58]
 Cuvette gap width 0.4 cm 0.4 cm [29, 45, 46]
 Volume of cells 600 μl 600 μl [26]
 Pulse parameters 2.0–2.5 kV; 25 μF; 200–800 Ω; 4–8 ms 2.0–2.5 kV; 25 μF; 200–800 Ω; 6–9 ms [44, 46]
Outgrowth    
 Recovery and plating medium YTG or 2×YTG 2×YTG [58, 59]
 Transformation efficiency Up to 106 transformants μg-1 DNA 2.4 × 101 transformants μg-1 pMTL85141 [35]
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