Electroporation entails brief, high intensity pulse to create transient pores in the cell membrane to facilitate the entry of exogenous macromolecules, which may otherwise be excluded. Removal of the external field leads to the resealing of the membrane electropores permitting the survival of the electrically stimulated recipient cells. Using this technique foreign deoxyribonucleic acid (DNA) has been successfully introduced into many cell types both from prokaryotes and eukaryotes. Increase in pulse voltage and length beyond a critical limit has been reported to decrease transformation efficiency, hence in this study we have investigated another strategy i.e. increase in the number of pulses at constant high voltage and pulse duration. Commonly used Agrobacterium strains LBA4404 and EHA101 and binary vector pCAMBIA1301 were used. Transformants were selected on a combination of hygromycin and kanamycin, and confirmed by polymerase chain reaction (PCR) and restriction analysis. Increase in the number of pulses was found to show a significant and linear increase in transformation efficiency.Application of deoxyribonucleic acid (DNA) technology to genetically manipulate cellular functions requires a suitable high efficiency transformation system. There are many approaches available for introducing foreign DNA into eukaryotic and prokaryotic cells, however electroporation is the most commonly used technique and currently of more practical interest. It is a gene transfection technique, which entails brief, high intensity pulse to facilitate the entry of exogenous molecules like DNA, ribonucleic acid (RNA), and protein. Removal of the external field leads to the resealing of the membrane electropores.Applying high intensity electric fields to reversibly permeablize biomembranes to create structural distortions, allowing the uptake of DNA was first demonstrated by Neuman in mouse myeloma cells (Neuman et al. 1982).
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