electroporation

MORRISJ at WSUVM1.CSC.WSU.EDU MORRISJ at WSUVM1.CSC.WSU.EDU
Sat Aug 29 23:52:20 EST 1992


On 28 Aug 92 10:39:50 GMT Neil Butt said:
>Dear networkers,
>We are about to embark on some plant transformation experimentation, using
>electroporation. All the protocols we have seen use 0.4ml cuvettes for
>plant cells. Our group has used electroporation techniques on yeast using
>0.2ml cuvettes. This may seem a petty factor, but we have not performed
>these experiments before and do not want to take any chances, but is it O.K
>to use the smaller sized cuvettes. We intend to do these experiments in both
>Tobacco and Arabidopsis in case the plant species is a factor in the
>protocol.
>Hope someone can help
>Neil Butt
>Biological Sciences
>University of Sussex
>Falmer
>Brighton
>East Sussex, BN1 9QG.
>U.K
>E-mail     bafp3 at uk.ac.sussex.central
>(from non-UK)   bafp3 at central.sussex.ac.uk
>
Dear Neil,

If you chose to use 0.2 cm cuvettes for electroporation, I'm confident that it
would work, but I'd suggest reducing the voltage setting by half.  Reducing
the cuvette width increases the field strength (volts per cm), and I'd
anticipate, reduces the pulse time.  I've had success with tobacco and
arabidopsis protoplasting in a thiosulfate buffer (Zaghmout et al. 1990, In
Vitro Cell. Dev. Biol. 26:315-317) and electroporating in a 0.45 M sucrose
buffer.  Pulse length seems to be important, I adjust cuvette volumes to get
an approximately 50-70 msec decay time.  Hope this helps.
John Morris  [morrisj at wsuvm1.csc.wsu.edu]



More information about the Arab-gen mailing list