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Are the Clonetics embryonic rat/mouse neuronal cells from a single donor or pooled?

The cells are pooled from whole litters, including both males and females.

Can I optimize the transfection efficiency or cell viability after the 4D-Nucleofector® Cell Line Optimization any further?

We offer a program fine tuning matrix for Optimization of Nucleofection® Conditions - A Short Guideline (lonza.com)Based on your current most satisfactory pulse(s) it provides guidelines which further programs to test to either increase cell...

What is the age of the rats and mice the Clonetics embryonic neuronal cells are isolated from?

Both are of embryonic origin: the rats are E18 or E19 and the mice are E14 or E15.


How many mycoplasma species exist? Which are the usual suspects in cell culture?

More than 180 species have been identified of which 20 distinct Mycoplasma and Acholeplasma species from human, bovine and swine have been isolated from cell culture. There are 6 species that account for 95% of all mycoplasma infections (M.orale,...

What are useful methods to enrich/purify specific hematopoetic cell populations before Nucleofection?

Specific blood cell populations can be enriched/purified for Nucleofection™ by MACS™ selection Kits (Miltenyi Biotec GmbH), RosetteSep™ Kits (StemCell Technologies Inc.) or Dynal™ beads (Invitrogen Corporation).

Do you recommend a purification method to purify specific blood/immune cell populations before Nucleofection®?

We recommend using negative selection (depletion) because your purified cells are "untouched", not influenced, and strictly not activated.Positive selection may cause increasing amounts of dead cells and could also lead to activation of the cells due...


DNA-purity and Nucleofection®: Can low A260:A280 ratios lead to both reduced transfection efficiency and cell viability?

Yes. To check the quality of your DNA we strongly recommend determining the A260:A280 ratio. It should be ideally 1.8 - 2.0 for a good DNA preparation, but least 1.6.

Can nicked DNA lead to reduced transfection efficiency in Nucleofection® experiments?

Yes. You should verify the integrity of your DNA on an agarose gel to see if it is degraded. Compare undigested plasmid to plasmid DNA digested with a suitable single cut restriction enzyme to linearize. Supercoiled plasmid will run faster than...
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