Isolation of a genetically accessible thermophilic xylan degrading bacterium from compost

Table 2 Overview of transformation optimisations for G. thermodenitrificans T12

Parameter changed	Medium	Growth^a (h)	Final OD^b	Voltage	Ω	µF	Cuvette width (mm)	Ptd^c (ms)	DNA^d (µg)	CFU/ µg DNA
N.A.	LB2 (2.50 g/L K₂HPO₄)	2	1.01	2000	200	25	2	3.7	3.031	4
Medium	LB2 (0.25 g/L K₂HPO₄)	1.67	0.96	2000	200	25	2	5.1	1.613	1240
Medium	LB2 (0.00 g/L K₂HPO₄)	1.66	0.96	2000	200	25	2	5	1.613	22
Wash buffer^e	LB2 (2.50 g/L K₂HPO₄)	1.67	0.96	2000	200	25	2	5.2	1.613	19
Medium/wash buffer^e	LB2 (0.00 g/L K₂HPO₄)	1.66	0.96	2000	200	25	2	5.4	1.613	2988
Medium/wash buffer^e	LB2 (0.25 g/L K₂HPO₄)	1.83	0.96	2000	200	25	2	5.5	1.564	17,071

^a Time of growth after dilution of the overnight pre-culture
^b Final OD₆₀₀ of the culture after growth prior to the washing step
^c Pulse time duration in milliseconds
^d Amount of plasmid (pNW33n) DNA added for transformation
^e Wash protocol was changed to 2 times washing with milliQ water (50 mL) followed by washing with 25 and 10 mL milliQ water +10 % glycerol

ISSN: 2731-3654