It is a bit difficult to predict accurately the effects of changing  
the gap width, but probably you can try the same set of conditions  
(voltage, capacity, temperature, etc.) if that gave satisfactory  
results with 0.1 cm cuvette. Pore formation efficiency seems more  
strongly dependent on the pulse width than field strength. Doubling  
the electrode gap will reduce the electric field by half and increase  
the resistance (and pulse width) four-fold, as long as the same  
volume of cell suspension is used (depth will be half), so each  
other's effects will be cancelled out to some extent.
Although low temperature slows pore resealing and is therefore not  
good for cell survival after pulses, it is believed that slow  
resealing is favourable for diffusion of the DNA into cells. In fact,  
electroporation works equally well at room temperature if lower  
voltage is used.
Kei Inouye

_____________________________
Kei Inouye
Department of Botany
Graduate School of Science
Kyoto University
Sakyo-ku, Kyoto 606-8502
Japan
_____________________________
telephone +81-75-753-4130
facsimile +81-75-753-4137
[log in to unmask]
http://cosmos.bot.kyoto-u.ac.jp/csm/
_____________________________


On 2009/03/04, at 10:35, Steve Charette wrote:

> Dear Dicty Colleagues,
>
> I need your help... We are trying to electroporate Dicty cells  
> using an old
> BioRad electroporator. We have 0.2 cm cuvette and we want to know  
> which
> parameters we should use to be equivalent to 0.85 kV and 25 µF when  
> using
> 0.1 cm cuvette.
>
> Another point, which is the best way to increase survival of the  
> cells: keep
> the cells on ice for 5 minutes after electric pulses or immediately
> resuspend the cells in HL5 after pulses?
>
> It will be great if someone can help us!
>
> Steve
>
> ---------------------------------------------------------
> ---------------------------------------------------------
> Steve Charette, Ph. D.
> Professeur adjoint (Dépt. Biochimie et microbiologie, U. Laval)
> Centre de recherche de l'Hôpital Laval
> Axe de recherche en pneumologie
> Pavillon Mallet (bureau M2689)
> Hôpital Laval
> 2725 Chemin Sainte-Foy
> Québec, Qc, Canada
> G1V 4G5
>
> Office: 418 656-8711 ext. 3311
> Laboratory: 418 656-8711 ext. 5871
> Fax : 418 656-4509
>
> WEBSITE: www.amibe.org
>
> [log in to unmask]
>