Q:

Following Nucleofection®, I cannot obtain stable clones from single cells. What could be the reason?

A:

Some cell lines need certain cell densities to proliferate.

For those cells you can try using conditioned medium for setting up limiting dilution or culturing single cells together with feeder cells.

Related Literature:
Lonza_BenchGuides_Guideline_for_Generation_of_Stable_Cell_Lines__Technical_Reference_Guide.pdf
Categories:
Transfection
Research Areas:
Gene Expression

You may also want to know:

Generation of a stable cell line: A positive control plasmid works, but I cannot obtain stable clones with my construct of interest. What could be the reason?
How long does it take to obtain stable transfectants?
What selection markers can I use for the generation of stable cell lines?
Is your pmaxGFP control vector supplied with the Nucleofector® Kits suitable for stable expression?
How many stable clones will I get from one transfection?
Why is the Nucleofector® Technology ideal for protein production from stable clones ?
In stable cell line generation, I have a very high transfection efficiency, but most of my cells die during selection. Is this to be expected?
How long is the MycoAlert® Mycoplasma Detection Assay signal stable?
What antibiotic concentration should I use for selecting stable transfectants?
How much DNA should I use in my Nucleofection® Reaction to obtain stable clones?
See More

Privacy | Legal | About Lonza

© Lonza. All rights reserved.