Journal: bioRxiv
Article Title: Active learning guides automated discovery of DNA delivery via electroporation for non-model microbes
doi: 10.1101/2025.11.18.689155
Figure Lengend Snippet: ( A-C ) Effect of buffer, voltage, and waveform on transformation efficiency (TE) in three Gram-negative bacteria. Data are the average of two biological replicates. ( A ) TE using four buffers: water, 25% sorbitol, 10% or 15% glycerol, and 25% sucrose. Data shown was electroporated at 3 kV using exponential decay (see additional voltages in Supplementary Figure 3). ( B ) TE using a range of electroporation voltages: 0.5-3 kV. Cells were washed with 10% glycerol ( E. coli ), 25% sorbitol ( S. amazonensis ), and 15% glycerol ( P. alcaliphila ), and electroporated using exponential decay waveform. Error bars represent standard error. ( C ) Comparison of TE using square or exponential decay waveforms. Data are results for all voltages and buffers tested in panel B. ( D ) Final parameter selection for 24-condition electroporation screen including four buffers, three voltages and two waveforms. ( E-F ) TE following the 24-condition electroporation screen performed on seven bacteria with a single plasmid using ( E ) exponential decay (EX) and ( F ) square (SQ) waveforms. Data are the average of two biological replicates, except P. sakaiensis and C. necator which are a single replicate.
Article Snippet: The plasmid was then transformed into dam–/dcm– electrocompetent E. coli (New England Biolabs) according to the manufacturer’s instructions, extracted and sequence verified by Eton Bioscience (Boston, MA).
Techniques: Transformation Assay, Bacteria, Electroporation, Comparison, Selection, Plasmid Preparation