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

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.

How many cortical neurons can I expect from a single pup?

According to QBM Cell Sciences, you can expect 12 million cortical neurons from an average E18-E19 rat pup. You can expect 4 million cortical neurons from an average mouse pup.

How much DNA should I use in my Nucleofection Reaction to obtain stable clones?

You should use the amount of DNA recommended in the optimized protocol for transient transfection. If obtaining a very high number of clones is important for your experiments, you can try increasing the amount of DNA. For further tips see our...

Does the 96-well Shuttle recognize if the cuvette plate is in backwards?

No, it does not recognize the orientation of the plate. Please make sure that well A1 is positioned in the upper left corner of the retainer.

Do any of the three instruments used in the 96-well Shuttle System "go to sleep" or power off automatically?

The energy options of the laptop are set as such that only the screensaver is activated, the harddisk won't turn down and hibernation is disabled. One can change these settings via the system control. However this is not recommended because by...

What is the molecular weight of your control pmaxGFP plasmid?

The molecular weight is approximately 2.3x10e6 g/mol based on the average MW of a base pair being 660 g/mol.

Is there a reason why in our Nucleofector Protocols for Primary Human Chondrocytes we include FCS in the pronase and collagenase solutions for preparation of chondrocytes?

Yes, even though this may sound odd since serum is generally known to inactivte proteases. We recommend it in this case, however, because the incubation times are quite long and the cells are stressed by the procedure. The amounts of enzymes should...

Does the GFP sequence in your pmaxGFP control plasmid have a stop codon?

Yes. The coding sequence ends with a stop codon (TGA/UGA).