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