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I currently work with some adherent cell lines, such as 293, which are grown in DMEM media. I have noticed that the mortality seems to be high after Nucleofection®. Do you have any recommendations?

...plate them in the appropriate media. There is no need to get rid of the RPMI, just add the cell suspension to the DMEM containing well. It is also possible to change the media after 5-6 hours.This will lower the mortality especially with 293 cells....
Missing: mycoalert

When I transfected my cells, error 8 appeared on the display of the Nucleofector® IIN, IIS or 2b. What is the reason?

...to the incomplete delivery; however, the cells can still be plated for analysis. This generally occurs because the conductivity of the solution was outside of accepted limits – this can happen if the media was not completely aspirated before resuspending, too much...
Missing: mycoalert

Can calcium influx issues affect viability or differentiation in Stem Cells after Nucleofection®?

When cells are transfected by Nucleofection®, transient pores are generated in the cell membrane. Generally, these pores disappear within 15 minutes of transfection, but if cells are plated immediately after Nucleofection® in medias...
Missing: mycoalert

Do you have any recommendation how to coat glass cover slips for neuron culture, e.g. for use with the Y unit or for post Nucleofection® culture in general?

...containing a glass coverslip). - Put back the lid onto the plate and incubate over night at room temperature (~22°C). - Wash twice with 1 ml of distilled water and let dry (open lid) under a sterile hood for approx. 2 hours.- Seed neurons If it generally...
Missing: mycoalert
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