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How much DNA can I use when transfecting hepatocytes with the standard Nucleofector?

We have used up to 6 µg of DNA per 100 µl reaction with no deleterious effect.

For hepatocytes, is plating density a concern post Nucleofection?

Yes, the optimized protocols for the standard Nucleofector™ recommend the use of 6 well plates. Plating into a 12-well plate will pose a problem as less cells will attach at high densities.

Four hours after Nucleofection, I can see the hepatocytes have attached to the well, but the morphology does not look correct, should I be concerned?

There is no reason for alarm. The hepatocytes may not exhibit normal morphology a few hours after Nucleofection™ but by 24 hours post Nucleofection™, morphology should be normal. Remember to perform a fluid change about 4 hours post Nucleofection™.

Do you have a protocol for the Nucleofection of Plasmodium berghei?

Yes. You can use our new Basic Parasite Nucleofector™ Starter kit for parasitic protozoa. The Basic Parasite Nucleofector Starter Kit (Cat.No. VMI-1001) should help you to determine the optimal program and Nucleofector™ Solution for your...

Do you have a protocol for the Nucleofection of Plasmodium yoelii?

Yes. You can use our new Basic Parasite Nucleofector™ Starter kit for parasitic protozoa. The Basic Parasite Nucleofector Starter Kit (Cat.No. VMI-1001) should help you to determine the optimal program and Nucleofector Solution for your...

Do you have a protocol for Nucleofection of parasites?

Yes. You can use our Basic Parasite Nucleofector™ Starter kit for parasitic protozoa. The Basic Parasite Nucleofector Starter Kit (Cat.No. VMI-1001) should help you to determine the optimal program and Nucleofector Solution for your...

What are the benefits of Amaxa Nucleofection over standard electroporation?

One major benefit of the Nucleofector™ Technology is that the nucleic acids are transferred directly into the nucleus enabling the transfection of non-dividing cells and an early analysis of transgene expression (depending on the protein) in as...

Is it more difficult to detect DsRed by FACS than GFP?

Yes, DsRed is much less bright than GFP and is more difficult to detect by FACS and microscopy because it bleaches quickly.

Is there a difference between, for example, the programs C-20 and C-020?

No, the programs C-20 and C-020 are identical regarding their electrical parameters. The letter minus a two-digit number is the nomenclature of the Nucleofector™ I. Since we have launched the Nucleofector™ II we changed it to a letter minus a three...

Why do you recommend 7AAD instead of PI staining for cell determination in mouse macrophages?

The results obtained using 7AAD staining are much clearer than those obtained using PI. Using flowcytrometry stained and unstained, transfected and non-transfected populations can be separated much more easily.
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