6533b857fe1ef96bd12b393c

RESEARCH PRODUCT

Transfer of plasmid DNA into cells with microelectroporation arrays on a chip

subject

description

The possibility to transfer pure DNA into bacterial cells forms the basis for the genetic engineering of the cell. Electroporation is a powerful and easy technique to introduce plasmid DNA into cells. Its drawback for use with high-throughput approaches is that with standard electroporation chambers the reactions have to carried out one after the other and the electroporation cuvettes are expensive. To obtain the possibility of high-throughput electroporation reactions Escherichia coli cells were electroinjected in parallel with different plasmids in reactions as small as 100 nl on a microstructured array of electrodes, forming hundred separate electroporation units on a chip of a square inch. The electroporation reactions reached about 1000 transformants per reaction. In addition microstructured electrode arrays in standard 96-, 384- and 1536-well microtiter plate format, which are compatible with robot arraying technology, were fabricated by means of photolithographically patterning and etching of photoresist coated copper films deposited on epoxy resin plates. Microelectroporation reactions were investigated with bacteria, and yeast.