2 replies to this topic

[metta]

hi,
I would like to know how important the time constant in electroporation of ES cells.

I'm using biorad gene pulser -II for electroporation. I use 240 volts and 500micro farady for electroporation. Every I get a time constant 8.2-8.8. Is this normal?

thanks

best
mm

[stemcellqueen]

I could be wrong, but I think the time constant can vary with the buffer you are electroporating cells in.

[vitalgene]

metta, on 07 October 2009 - 06:06 AM, said:

hi,
I would like to know how important the time constant in electroporation of ES cells.

I'm using biorad gene pulser -II for electroporation. I use 240 volts and 500micro farady for electroporation. Every I get a time constant 8.2-8.8. Is this normal?

thanks

best
mm

tc of approx 9 or more is give best results on clone generation
with the condtions mentioned above , according to me its a good electroporation and shld lead optimum trasfectiong efficeincy
i use PBS devoid of CaMg ions, and use bioradcuvett 4mm with 10ug pDNA, works almost always
the time constast is factor of time taken for electoric impluse to  travel between the 2plate, meaning the cells. since the medium is devoid of ion so the only way of electric conductance is via cells acting a conductor of electricityy and hence the dna

hope this is helpful
good luck
vg