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Are there positive controls for the MycoAlert Assay?

The MycoAlert™ Assay Control Set (LT07-518) is available separately and includes a lyophilized positive control (1 ml) and assay buffer (2 ml) for reconstitution. The assay buffer also serves as a negative control. The positive and negative controls...

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

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

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.

Does the MycoAlert Assay recognize the whole spectrum of mycoplasma?

The mycoplasma specific metabolism detected by the MycoAlert™ Assay is present in all members of the mollicute family (Mycoplasma, Acholeplasma, Entomoplasma and Spiroplasma) except Ureaplasma (which are no usual suspects in cell culture). 95% of all...

Can nicked DNA lead to reduced transfection efficiency?

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...

What antibodies can be used against the GFP produced by pmaxGFP and pmaxFP-Green vectors? What antibodies can be used to detect denatured GFP? What antibodies can be used to detect the RFP expressed by pmaxFP-Red?

Most of the commercial antibodies for GFP do not detect GFP expressed from pmaxGFP or pmaxFP-Green vectors. The origin of the protein expressed by the pmaxGFP control plasmid and pmaxFP-Green is from the green fluorescent protein CopGFP, cloned from...

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 characterized...

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

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 perform an...

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. We have demonstrated that the MycoAlert™ Kit will detect mycoplasma...

What is the shelf life of the 96-well Shuttle kits?

The shelf life of the 96-well Shuttle™ Kits is 24 months from date of manufacturing. We guarantee a remaining shelf life of at least 6 months after sending them out of the warehouse.
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