Chemistry: molecular biology and microbiology – Apparatus – Including measuring or testing
Patent
1990-07-09
1992-03-24
Niebling, John
Chemistry: molecular biology and microbiology
Apparatus
Including measuring or testing
435173, 4351721, 935 52, 935 85, C12N 1300, C12N 1500
Patent
active
050988433
ABSTRACT:
Apparatus and methodology for the transformation of living cells by the introduction of DNA thereto through transient pores created in the cell walls by the highly efficient electroporation of such cells in a liquid suspension with DNA. High efficiency electroporation is accomplished by the generation of a high voltage electrical pulse with precise characteristics determined necessary to optimize cell transformation and the application of such pulse to a sample of such suspension or by continuous flow of the suspension through a static electrical field under flow conditions to simulate the precise characteristics of such high voltage pulse. Thus, the suspension, whether treated as a non-flowing sample or as a continuous flow liquid suspension of cells in admixture with DNA, is subjected to a high intensity electric field at its maximum intensity in no more than about 100 microseconds and thereafter to reduced field intensity to zero field intensity in no more than about 5,000 microseconds. The high intensity pulse, or static electrical field with suspension flow therethrough simulating the characteristics of such pulse, porates the wall of the cells in suspension thereby allowing the introduction of DNA into the cells with a high survival rate for such cells.
REFERENCES:
patent: 4561961 (1985-12-01), Hofmann
patent: 4699881 (1987-10-01), Matschke
patent: 4750100 (1988-06-01), Ragsdale
Junkins Philip D.
Niebling John
Starsiak Jr. John S.
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