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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 × 105 ) at room temperature. Escherichia coli TOP10 cells and plasmid DNA are sequentially injected into a 50 μm 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%.

Item Type:	Refereed Article
Keywords:	molecular biology, microbiology
Research Division:	Chemical Sciences
Research Group:	Analytical chemistry
Research Field:	Separation science
Objective Division:	Expanding Knowledge
Objective Group:	Expanding knowledge
Objective Field:	Expanding knowledge in the biological sciences
UTAS Author:	Alves, MN (Ms Monica Alves)
UTAS Author:	Powell, SM (Dr Shane Powell)
UTAS Author:	Macka, M (Professor Mirek Macka)
UTAS Author:	Breadmore, MC (Professor Michael Breadmore)
ID Code:	154717
Year Published:	2022
Deposited By:	Agriculture and Food Systems
Deposited On:	2022-12-23
Last Modified:	2023-01-10
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