Electroporation 2-mm or 4-mm?

March 22, 2011 at 6:59 am #14703

Participant

Hello.
I am trying to perform electroporation on many kinds of mouse cell-lines by using Bio-rad electroporator.
I can find protocol here and there such as literatures and Bio-rad websites.
However it seems that there are many different kinds of protocols for even a single cell line.
What is more confusing me is the uses of 2-mm and 4-mm cuvettes.

I thought that 2-mm is for fungus and 4-mm is for mammalian cells.
But some protocols use 2-mm and some uses 4-mm for the same kind of cells, of course with different parameters.

Is there a formula I can use to convert parametesr for 4-mm cuvette to 2-mm cuvette?

Thank you.

April 12, 2011 at 12:32 am #104405

AntonAleraLabs

Participant

Hi Evilid – It is quite easy to compare protocols actually: you always need to operate in terms of field intensity. Field intensity=Voltage/Gap size of your cuvette in cm (0.2 or 0.4 in your case. For mouse cell lines aim for 500 V/cm or 100V for 0.2 cm cuvette and 200V for 0.4 cm cuvette. This is a good start. Use 500 uF or 960 uF capacitance setting for mammalian cell lines.

Good luck,

Hi Tom – Vitamin D does NOT work well for THP-1. You need to use Retinoic Acid at 10 nM or more for 72 h to differentiate THP-1 cells. Large proportion of cells (up to 90%) will die if the protocol is working. Cell will be phagocytic for 3 u latex beads.

Another option is Mono Mac 6 cell line which is phagocytic right out of the tube. I have not tried it because license to use it is quite expensive for a company, but for academia it is free from the German Collection of Microorganisms and Cell Cultures:
http://www.dsmz.de/

Good luck, feel free to contact me if you have any questions.

—
Anton
Principal Consultant, Molecular and Cell Biology, Biomarkers

Phone: (518) 334-0922
E-mail: antonb@aleralabs.com
Web: http://www.aleralabs.com

Alera Labs
P.O. Box 13019
Research Triangle Park, NC 27709

August 4, 2011 at 12:07 pm #105826

Gomer321

Participant

I can’t find any information about Electroporation. Give me some useful links where I can learn throughly about it.

Thanks in advance!!!

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