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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.

When performing the MycoAlert® Mycoplasma Detection Assay, why is it necessary to spin out the cells?

It is highly recommended that the samples are spun down prior to testing. If cells are present in the assay they will contribute to the initial reading producing a higher background. This in turn can decrease the difference between the background and...

How long is the MycoAlert® Mycoplasma Detection Assay signal stable?

Please follow the protocol exactly as written – add reagent, wait 5 min and read, add substrate, wait 10 minutes and read again. 

Is the MycoAlert® Assay validated for use in GMP environment?

No. The MycoAlert® Assay is sold for Research Use Only.

In my Nucleofection Experiment I see lower expression with my IRES construct in comparison with you pmax-GFP control. Do you have any information about the different expression profiles?

The original attenuation of the IRES sequence was performed to allow for a greater difference between the expression levels of the upstream gene of interest and the downstream reporter gene. If the downstream reporter gene, the product of less than...

Can I also use the Lucetta® 2 Luminometer for other assays besides the MycoAlert® Mycoplasma Detection Assay?

Yes. In addition to the tailor-made MycoAlert® Mode it also has a "Single Read" mode which is suited for unprocessed luminescence readings, e.g. ATP-based cell proliferation or cytotoxicity assays, or Luciferase reporter gene assays.

I thawed and plated my Clonetics embryonic rat or mouse neuronal cells and there are a lot of dead cells. What happened?

Cell death will usually be observed during the first few days of growth, resulting in cell debris in the culture – this is normal. Cell cultivation should be continued and surviving cells should start to develop. By day 4, neurite networks will be...

What are the best time points for analysis of my siRNA experiments?

As the stability and half-life of various mRNAs and their protein products varies, it is important to empirically determine the best time points for assessing target knockdown. For example, it has been documented that in mammalian cells, mRNA...

Can I co-transfect oligonucleotides and plasmids with the Nucleofector® Technology?

Yes. As the same protocol applies for any nucleic acid substrate (vectors or oligonucleotides) you can easily perform co-transfections either for transfection control or rescue experiments.

What type of matrix should your Clonetics embryonic rat/mouse neural cells be seeded onto?

For optimal results, poly-D-lysine with laminin should be used, especially for the hippocampus cells.Poly-D-lysine without the laminin or poly-L-lysine could also be used for the rat and mouse neuronal cells.
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