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How often do mycoplasma contaminations occur in cell cultures?

A conservative estimate states that between 15-35% of all continuous cell cultures are contaminated with mycoplasma (Drexler HG, Uphof CC; 2002), some estimates are even higher (up to 80 % in some countries (Koshimizu K, Kotani H; 1981).

What is the ratio of Supplement to Nucleofector® Solution?

The ratio is 1:4,5 (500 µL of supplement is added to 2.25 mL of Nucleofector® Solution).For a single 100 µL reaction, use 18 µL of supplement plus 82 µL of solution to make the 100 µL.

Can I use SDS as a detergent for a positive control if I do cytotoxicity assay with ToxiLight?

No. SDS has a negative influence on Adenylate Kinase (AK) activity. SDS is a cationic detergent, it is a very powerful protein denaturing agent, you will be aware of its use in SDS-PAGE for those reasons. ToxiLight™ works by measuring the activity of...

What is the molecular weight (MW) of your control plasmid pmaxGFP?

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

Which luminometer is suited for performing the MycoAlert® Assay?

For performing the MycoAlert® Assay in theory every luminometer is suitable as long as a specific sensitivity is given. Please find attached our list with luminometers we know to be suited to run the MycoAlert® and MycoAlert® Plus Mycoplasma...


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...

What are useful methods to enrich/purify specific hematopoetic cell populations before Nucleofection?

Specific blood cell populations can be enriched/purified for Nucleofection™ by MACS™ selection Kits (Miltenyi Biotec GmbH), RosetteSep™ Kits (StemCell Technologies Inc.) or Dynal™ beads (Invitrogen Corporation).

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...

Do you recommend a purification method to purify specific blood/immune cell populations before Nucleofection®?

We recommend using negative selection (depletion) because your purified cells are "untouched", not influenced, and strictly not activated.Positive selection may cause increasing amounts of dead cells and could also lead to activation of the cells due...
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