Mycoplasmacheck Service -
Mycoplasma testing in every cell line

Details of our Mycoplasmacheck

Full-service package offered by Mycoplasmacheck:

• Detection of all mycoplasma species listed in the European Pharmacopoeia 2.6.7: M. arginini, M. fermentans, M. orale, M. hyorhinis, M. hominis, M. genitalium, M. salivarium, M. synoviae, M. pirum, M. gallisepticum, M. pneumoniae, M. yeatsii, Spiroplasma citri and Acholeplasma laidlawii
• Identification of more than 100 additional mollicutes
• Highly sensitive assay capable of detecting 1 international unit (IU) per PCR of the World Health Organization’s (WHO) international standard for mycoplasma DNA (Please see application note)
• Highly specific test that identifies ten copies of mycoplasma per reaction with no cross-reactivity to E. coli or eukaryotic DNA
• Use of positive and negative controls to monitor test performance
• Exclusively licensed from the German Cancer Research Center, (Deutsches Krebsforschungszentrum, DKFZ), Heidelberg, Germany
• Easy sample drop-off at any Collection Point or ship separately

Technology used for Mycoplasmacheck

• Roche LightCycler 480 II

Deliverables of Mycoplasmacheck

• Data report indicating mycoplasma status, as well as any PCR inhibitors found in the sample
• The user can compile a PDF document online at any time to view the results from any specific testing period.
• PDF document serving as an independent certificate of results; suitable for direct submission to journals or granting agencies

Reliably detect all common mycoplasma species

Mycoplasma contamination adversely affects 35% of the world’s cell lines.

As signs of mycoplasma are easy to miss, only regular monitoring of cell cultures can ensure that research results are valid and reproducible. With its standardised workflows, we can rapidly and reliably detect all common mycoplasma species.

Table 1. List of mycoplasma species frequently detected in cell cultures. These mycoplasma, all found in the European Pharmacopoeia 2.6.7., are readily detectable with Mycoplasmacheck.

As a qPCR-based assay, the technique is more accurate and sensitive than traditional microbial culture methods and bioluminescence-based methods for mycoplasma detection (Fig1). Besides severely reducing the hands-on time involved in in-house testing, outsourcing the task of mycoplasma detection offers distinct benefits that cannot be matched by do-it-yourself kits (Fig 2).

Fig. 1. Comparison of three different methods for detection of mycoplasma in cell cultures. The graph demonstrates the superiority of mycoplasma PCR over culture-based or bioluminescence-based (Mycoalert) techniques in analysing cell lines for mycoplasma (adapted from Cytotechnology (2014) 66:861–873)

Fig. 2. Advantages of using Mycoplasmacheck for mycoplasma PCR detection over in-house PCR kits

Here you can find a selection of publications from our customers, that tested their cell lines for mycoplasma contamination

2022

Mycoplasmacheck: Gizon et. al., Generation of a heterozygous SCN5A knockout human induced pluripotent stem cell line by CRISPR/Cas9 edition

2021

Mycoplasmacheck: Kristiansson et. al., 177Lu-PSMA-617 Therapy in Mice, with or without the Antioxidant α1-Microglobulin (A1M), Including Kidney Damage Assessment Using 99mTc-MAG3 Imaging

Mycoplasmacheck: Goisnard et. al., The New Serum-Free OptiPASS® Medium in Cold and Oxygen-Free Conditions: An Innovative Conservation Method for the Preservation of MDA-MB-231 Triple Negative Breast Cancer Spheroids

Mycoplasmacheck: Sacchi et. al., Nanogold Functionalized With Lipoamide-isoDGR: A Simple, Robust and Versatile Nanosystem for αvβ3-Integrin Targeting 

Mycoplasmacheck: Malakpour-Permlid et. al., Identification of extracellular matrix proteins secreted by human dermal fibroblasts cultured in 3D electrospun scaffolds

Mycoplasmacheck: Groisnard et. al., LightSpot®-FL-1 Fluorescent Probe: An Innovative Tool for Cancer Drug Resistance Analysis by Direct Detection and Quantification of the P-glycoprotein (P-gp) on Monolayer Culture and Spheroid Triple Negative Breast Cancer Models

Mycoplasmacheck: Boehm et. al., SMG5-SMG7 authorize nonsense-mediated mRNA decay by enabling SMG6 endonucleolytic activity

Mycoplasmacheck: Perrin-Cocon et. al., A hexokinase isoenzyme switch in human liver cancer cells promotes lipogenesis and enhances innate immunity  

Mycoplasmacheck: Plavec et. al., Targeting of fluorescent Lactococcus lactis to colorectal cancer cells through surface display of tumour-antigen binding proteins

2020

Mycoplasmacheck: Janz et. al., Generation of two patient-derived iPSC lines from siblings (LIBUCi001-A and LIBUCi002-A) and a genetically modified iPSC line (JMUi001-A-1) to mimic dilated cardiomyopathy with ataxia (DCMA) caused by a homozygous DNAJC19 mutation 

Mycoplasmacheck: Nesteruk et. al., Forced expression of HOXA13 confers oncogenic hallmarks to esophageal keratinocytes

Mycoplasmacheck: Gretarsson et. al., Dppa2 and Dppa4 counteract de novo methylation to establish a permissive epigenome for development

Mycoplasmacheck: Tesauro et. al., Different Camptothecin Sensitivities in Subpopulations of Colon Cancer Cells Correlate with Expression of Different Phospho-Isoforms of Topoisomerase I with Different Activities

Mycoplasmacheck: Chua et. al., The NALCN channel complex is voltage sensitive and directly modulated by extracellular calcium

Pre-requisites prior testing

For maximum sensitivity: Recommendations for cell culture cultivation prior to testing:

• Cell lines should be cultured in the absence of mycoplasma-active antibiotics for one week prior to submission. Penicillin and streptomycin do not affect mycoplasma nor do they inhibit MYCOPLASMACHECK.
• Cell cultures should be maintained in the same media without dilution with fresh media for three days before testing.
• Cryopreserved cell cultures should be cultured about two weeks prior to sample submission.
• 80-90% confluence of the tissue culture is recommended for testing. Inhibiting substances may accumulate in >90% confluent cultures.

Sample preparation

Sending samples

Sending samples according to the requirements below helps us to do our job better
and provides you with accurate results.

 Shipping address (if no dropbox is used):

Eurofins Genomics Europe Applied Genomics GmbH, Christian Dorn, Building 5,

Anzinger Str. 11, 85560 Ebersberg, Germany

Sample submission

• Use 1.5 ml safe-lock tubes for your templates and primers
• Do not tape or wrap tubes with parafilm. Safe-lock tubes offer perfect sealing and evaporation protection
• Label your template with our Mycoplasmacheck barcode labels.
  
• Use a water resistant marker for any additional labeling of your tubes

This picture helps you to stick the labels correctly to the tube. Please note, these are NOT MPC barcodes. But still it should be handled the same way.

Mycoplasmacheck (MPC) Barcodes

• The Barcodes can be found in the "Barcode Management Genotyping" list

Order updates

• When your samples arrive in our lab an Email will be sent to you and an order will be visible under my orders
• When samples are finished you will receive a "Partially complete" or a "Complete" Email
  • Partially complete = not all samples are finished
  • Complete = all samples are finished
• The results are available in the Mycoplasmacheck results
  • You can easily create a report for one or more samples
  • Share your results with your colleagues
  • Sort and filter according to the results and date range