Amaxa Nucleofector vs Neon thermofisher electroporator systems?

Our lab 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 primary myoblasts, any opinions/preferences?

In the manual they recommend use the tips by only two times, I and using from 3 times, does anyone using for more times with good results?

Recently, we started using the Neon Transfection System in our lab. However, a very confusing concern popped up, and I would really appreciate your kind advice a lot. It is about the decision to use either the 10 or the 100 uL tip. For example, I am now planning for a transfection experiment using siRNA in order to have cell pellets afterwards for q-RT-PCR and WB. So I suppose the 100 uL tip which allows transfection of about 0.5 to 2 million cells would be more suitable. However, I also noticed that the preset protocols on the Thermo Neon website for HCT-116 or HT-29 or several other cells are more commonly specified using 10 uL tip !! So in such case, would it be possible to use the same stated voltage and pulses but for using 100 uL tip ?????? Many thanks in advance,

I am working with new fish cell lines especially from cold-water fishes, so how can we choose the best program and process to get cells with higher viability and transfection efficiency? Suggestions from experiences using the Neon® Transfection System for transfection with Plasmid, Protein, and RNP.-complex are highly appreciated.

Electrolyte buffer E is for 10ul transfection kit.

Electrolyte buffer E2 is for 100ul transfection kit.

Just wondering if these two can be interchangeable as I have finished the buffer E but still have some E2 buffer left.

Thanks.

I am working with iPSC cell culture and finding contamination after two or three days. Media color changes yellowish green and turbid. I was not able to see any cells after the contamination even after 70-80%confluency. I am attaching pictures of the plate.

Hello everyone,

I am trying to make a CRISPR-Cas9 knockout on lung cancer primary cells using the RNP complex. I am using the Neon Transfection System but so far it failed - all the cells that took the complex were dead. The settings I tried were: puls voltage 1200 v, width 30 ms, pulse number 2.

I am planning to use their optimization protocol on the 24-well plate but maybe any of you have already tried a similar edition on any type of primary cancer cells and can share the settings used?

Hi,

I need to plot MFI for different channels I used when FACS-characterizing P0-P4 cells. However, I am confused on how to do so. First, I basically selected in "Statistics" in Flow Jo the option "Median" to get MFI for the channels I'm interested in but im not sure if the value is given in % since some values are around 13.9 while others are approximately 124, for example. In addition, I am not sure on how should I organize or plot this data since I have never done this before. If someone could help me, I'd be grateful!

Dear everyone,

May you please recommend a protocol for electroporation of NK-92 cells using Neon machine? I have never done that before and woul dbe happy for recommendations if anyone had experience with them.

There is recommendation for NK-92 cell electroporation, but any suggestions are welcome.

Thanks,

Kind regards,

Maria