Electroporation : Nucleofector (Amaxa) vs Neon (LifeTechnology) Systems

[Dars]

My group is looking into purchasing an electroporator and we are in the middle of demoing the Nucleofector (Amaxa) vs Neon (LifeTechnology) systems. Does anyone have experience with using these two for electroporating CRISPR plasmids, any opinions/preferences?

We are mainly working with human primary fibroblasts and myoblasts and have had disappointing success with lipofectamin 3000

Thank you!

[Elph]

Hi Dars,

I have used both machines extensively with mouse embryonic stem cells. Both work great in terms of efficiency (>80% of green cells with the pmaxGFP reporter).

Neon is nice because you only buy one kit and you only change the electroporation settings between cell lines, whereas with Amaxa you need different kits for different cell lines. And it is much cheaper. If anything II found that viability is less good than with Amaxa (for mouse ES cells) but it not too bad. And I find that the R solution in the Neon kit goes bad after a couple of months but if your turnover is faster that should not be a problem.

For these reasons if I was to buy a machine I would pick the Neon system but you should definitely ask for a demo to test both machines on your cells before you make any decision.

Switching to these electroporators from lipofectamine was night and day for my genome editing.

Good luck with your experiments!

Elphege

[Ran Antes]

Hi Elphege.

Did you try tranfecting the mESC with lipofectamine?

do you think nucleofection is better for this type of cells?

could you elaborate about the amount of DNA used in this method?

 I was trying to transfect with lipo3000, but with no success, and would really appreciate any help :-)

Thanks,

Ran

[Ran Antes]

HI Elphege.

I have the amaxa nucleofector in the lab, and the thing is that it has 4 different programs for mESC. I tries 2 of them (A23; A24) but it killed most of my cells.

Can you recommend a specific program?

Im trying to transfect my gRNA and a ssODN and I get very low efficiency.

Appreciate any help.

Thanks,

Ran

[Phil Abbosh]

Ran
I had some trouble using the amaxa machine and the optimization kit on one particular cell line and called the company (lonza). they were able to recommend some other programs and one worked pretty well (50% GFP+ with little cell death).  you might try calling.

phil

[JJ]

Clontech's Xfect works nicely for us in mES cells. We no longer use electroporation for transfection. Low passage number too.