e coli c2925  (New England Biolabs)

Journal: Microbial Biotechnology

Article Title: Developing High‐Efficiency Electroporation Protocols for Hard‐To‐Transform Halomonas spp.

doi: 10.1111/1751-7915.70285

Figure Lengend Snippet: Effect of plasmid source on the electroporation efficiency of Halomonas elongata DSM 2581. (a) Electroporation efficiencies of pSEVA241 purified from either E. coli 10‐beta (NEB) or H. elongata DSM 2581. (b) Electroporation efficiencies of pSEVA231 purified from either E. coli 10‐beta (NEB), E. coli C2925 (NEB) or H. elongata DSM 2581. Data shown represent mean ± standard deviation from three biological replicates. Negative control experiments were performed by electroporating electrocompetent cells without the addition of plasmid pSEVA241 (a) or pSEVA231 (b) (* p < 0.05; *** p < 0.001; **** p < 0.0001).

Article Snippet: Escherichia coli 10‐beta and E. coli C2925 ( dam − , dcm − ) were sourced from New England Biolabs (NEB).

Techniques: Plasmid Preparation, Electroporation, Purification, Standard Deviation, Negative Control

Journal: Microbial Biotechnology

Article Title: Developing High‐Efficiency Electroporation Protocols for Hard‐To‐Transform Halomonas spp.

doi: 10.1111/1751-7915.70285

Figure Lengend Snippet: Electroporation of Halomonas boliviensis LC1 and Halomonas campaniensis LS21. Electroporation efficiencies of H. boliviensis LC1 and H. campaniensis LS21 transformed with pSEVA231 purified from either E. coli C2925 (NEB) (a) Comparison of electroporation efficiencies from electrocompetent cells prepared from cultures grown in LB medium containing different concentrations of NaCl: 6% vs. 1%. (b) Comparison of electroporation efficiencies using two different electroporator systems—Bio‐Rad MicroPulser vs. Bio‐Rad Gene Pulser—under varying electroporation conditions: Voltage, pulse number and resistance. Parameters for conditions C1, C4, C6 and C7 are detailed in Figure . Data shown represent mean ± standard deviation from three biological replicates. Negative control experiments were performed by electroporating electrocompetent cells without the addition of plasmid pSEVA231. (*** p < 0.001; **** p < 0.0001)

Article Snippet: Escherichia coli 10‐beta and E. coli C2925 ( dam − , dcm − ) were sourced from New England Biolabs (NEB).

Techniques: Electroporation, Transformation Assay, Purification, Comparison, Standard Deviation, Negative Control, Plasmid Preparation

Journal: Microbial Biotechnology

Article Title: Developing High‐Efficiency Electroporation Protocols for Hard‐To‐Transform Halomonas spp.

doi: 10.1111/1751-7915.70285

Figure Lengend Snippet: Effect of plasmid source on the electroporation efficiency of Halomonas elongata DSM 2581. (a) Electroporation efficiencies of pSEVA241 purified from either E. coli 10‐beta (NEB) or H. elongata DSM 2581. (b) Electroporation efficiencies of pSEVA231 purified from either E. coli 10‐beta (NEB), E. coli C2925 (NEB) or H. elongata DSM 2581. Data shown represent mean ± standard deviation from three biological replicates. Negative control experiments were performed by electroporating electrocompetent cells without the addition of plasmid pSEVA241 (a) or pSEVA231 (b) (* p < 0.05; *** p < 0.001; **** p < 0.0001).

Article Snippet: Following this approach, we tested electroporating H. elongata with plasmids purified from the dam − / dcm − E. coli C2925 (NEB).

Techniques: Plasmid Preparation, Electroporation, Purification, Standard Deviation, Negative Control

Journal: Microbial Biotechnology

Article Title: Developing High‐Efficiency Electroporation Protocols for Hard‐To‐Transform Halomonas spp.

doi: 10.1111/1751-7915.70285

Figure Lengend Snippet: Electroporation of Halomonas boliviensis LC1 and Halomonas campaniensis LS21. Electroporation efficiencies of H. boliviensis LC1 and H. campaniensis LS21 transformed with pSEVA231 purified from either E. coli C2925 (NEB) (a) Comparison of electroporation efficiencies from electrocompetent cells prepared from cultures grown in LB medium containing different concentrations of NaCl: 6% vs. 1%. (b) Comparison of electroporation efficiencies using two different electroporator systems—Bio‐Rad MicroPulser vs. Bio‐Rad Gene Pulser—under varying electroporation conditions: Voltage, pulse number and resistance. Parameters for conditions C1, C4, C6 and C7 are detailed in Figure . Data shown represent mean ± standard deviation from three biological replicates. Negative control experiments were performed by electroporating electrocompetent cells without the addition of plasmid pSEVA231. (*** p < 0.001; **** p < 0.0001)

Article Snippet: Following this approach, we tested electroporating H. elongata with plasmids purified from the dam − / dcm − E. coli C2925 (NEB).

Techniques: Electroporation, Transformation Assay, Purification, Comparison, Standard Deviation, Negative Control, Plasmid Preparation

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: Individual pGL2 plasmids ( Supplementary Table 1 ) were stored in TransforMax EC100D pir-116 cells (Biosearch Technologies); pAKgfp1-kan was stored in dam–/dcm– E. coli (New England Biolabs).

Techniques: Transformation Assay, Bacteria, Electroporation, Comparison, Selection, Plasmid Preparation

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