Detail publikačního výsledku

Isotachophoresis for rapid transformation of Escherichia coli

ALVES, M.; NAI, Y.H.; POWELL, S.M.; MACKA, M.; BREADMORE, M.

Originální název

Isotachophoresis for rapid transformation of Escherichia coli

Anglický název

Isotachophoresis for rapid transformation of Escherichia coli

Druh

Článek WoS

Originální abstrakt

A frequent limitation of electroporation (EP) and chemical transformation (CT) are the need of tedious and time-consuming procedures for inducing transformation competence, the substantial number of cells required, and the low transformation yields typically achieved. Here, we show a new and rapid electrokinetic method for transformation of small number of noncompetent Escherichia coli TOP10 cells (2-3 x 10(5)) at room temperature. Escherichia coli TOP10 cells and plasmid DNA are sequentially injected into a 50 mu m ID capillary and focused into 11.5 nL by isotachophoresis (ITP) induced by application of high DC voltage (-16 kV). Through ITP, a large excess of plasmid DNA is brought in contact with the cell surface, with the contact time adjusted by application of a counter-pressure (1.3 psi) opposing the ITP movement. The transformation rate was more than 1000-fold higher compared to EP and CT at survival rates greater than 60%.

Anglický abstrakt

A frequent limitation of electroporation (EP) and chemical transformation (CT) are the need of tedious and time-consuming procedures for inducing transformation competence, the substantial number of cells required, and the low transformation yields typically achieved. Here, we show a new and rapid electrokinetic method for transformation of small number of noncompetent Escherichia coli TOP10 cells (2-3 x 10(5)) at room temperature. Escherichia coli TOP10 cells and plasmid DNA are sequentially injected into a 50 mu m ID capillary and focused into 11.5 nL by isotachophoresis (ITP) induced by application of high DC voltage (-16 kV). Through ITP, a large excess of plasmid DNA is brought in contact with the cell surface, with the contact time adjusted by application of a counter-pressure (1.3 psi) opposing the ITP movement. The transformation rate was more than 1000-fold higher compared to EP and CT at survival rates greater than 60%.

Klíčová slova

Bacteria; CE; Isotachophoresis; Plasmid; Transformation

Klíčová slova v angličtině

Bacteria; CE; Isotachophoresis; Plasmid; Transformation

Autoři

ALVES, M.; NAI, Y.H.; POWELL, S.M.; MACKA, M.; BREADMORE, M.

Rok RIV

2022

Vydáno

24.02.2022

ISSN

0173-0835

Periodikum

ELECTROPHORESIS

Svazek

43

Číslo

4

Stát

Spolková republika Německo

Strany od

543

Strany do

547

Strany počet

5

URL

https://pubmed.ncbi.nlm.nih.gov/34837243/

BibTex

@article{BUT176764,
  author="ALVES, M. and NAI, Y.H. and POWELL, S.M. and MACKA, M. and BREADMORE, M.",
  title="Isotachophoresis for rapid transformation of Escherichia coli",
  journal="ELECTROPHORESIS",
  year="2022",
  volume="43",
  number="4",
  pages="543--547",
  doi="10.1002/elps.202100095",
  issn="0173-0835",
  url="https://pubmed.ncbi.nlm.nih.gov/34837243/"
}