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6402 results sorted by

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.

Do bacteria lead to false positive MycoAlert® Results?

NO - the lysis reagent in the MycoAlert® Assay is not strong enough to lyse bacteria / yeast. Also, the enzymes utilized are not very common in bacteria or yeast – but are universal among the mollicute family. Bacterial contamination is...

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...

Can mycoplasma grow in the absence of cells?

There is evidence to suggest that mycoplasma can survive in the absence of cells, some may even proliferate, but as a rule they are much more viable if cells are present. 

What protocol should I use for Nucleofection® of patient derived blood samples, e.g. leukemia or lymphoma cells?

Unfortunately, we do not have a ready-to-use protocol for Chronic lymphocytic leukemia (CLL) or other blood cell derived cancer cells. The cells often have a chromosomal aberration and show the properties of a cell line. Therefore we suggest to...

Is it possible to store reconstituted MycoAlert®/MycoAlert® PLUS Reagent and Substrate?

For optimal assay conditions, reconstituted reagent and substrate should be used fresh within 2 hours after reconstitution.During this time they can be kept at room temperature.For the MycoAlert® Kit: Unused, reconstituted components can be aliquoted...

How much sample do you need for accurate results with MycoAlert® Assay? Do you need cells or just supernatant?

The standard protocol calls for taking 2 ml of culture media, spinning down any cells and removing 100 µl of supernatant as sample.Smaller initial aliquots of media may be taken to start with if necessary.

My suspension-adapted HEK293 (or CHO) cells tend to clump in culture. How can I avoid this?

Efficient protein production experiments require growth of the host cell in single-cell suspension, something that is sometimes difficult to achieve since most established cell lines retain their adherent growth characteristics. Commercially...

Is it possible to quantify the mycoplasma with the MycoAlert® Mycoplasma Detection Assay?

No, the MycoAlert® Mycoplasma Detection Assay (as well as the MycoAlert® PLUS Mycoplasma Detection Assay) is not quantitative; it has been designed to give a simple Yes or No answer, pertaining to the presence of Mycoplasma.

How can I improve Human Hepatocyte viability post Nucleofection on the 96-well Shuttle device?

In addition to proper cell handling and using culture media optimized for long term culturing of hepatocytes (Lonza Basal medium supplemented with BulletKit), we recommend separating the healthy hepatocytes from the dead cells and debris in the...
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