J4-4324 — Annual report 2011
1.
Analysis of gene electrotransfer and electropermeabilization for sub-optimal plasmid concentrations

In previous studies we have shown that low concentartions are relevant for the in vivo conditions (in tissues). The interaction of DNA with the membrane in vitro at different pulsing protocols was observed with intercalation dye TOTO. We found that for introduction of small molecules high voltage (HV) pulse is sufficient, while for more efficient introduction of plasmid DNA into cell requires a combination of HV and LV pulses, where LV-pulse represents the electrophoretic pull of DNA to the cells and facilitate transport through the membrane, while the HV-pulse is responsible for cell membrane electroporation. For the quality and quantity analysis of electropermeailization we also compared two methods (spectrofluorimetric and microscopic analysis). We found that the spectrofluorimetric method is suitable for qualitative and quantitative analysis, while microscopic analysis is quantitative method and less accurate.

F.01 Acquisition of new practical knowledge, information and skills

COBISS.SI-ID: 8878420