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        Mycoplasmacheck

    Mycoplasmacheck Service -
    Mycoplasma testing in every cell line

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    • Mycoplasmacheck
     

     Easy outsourcing of mycoplasma testing in any cell line

     


    Mycoplasmacheck is a qPCR-based service for detection of mycoplasma in cell cultures.

     

    >> Request Free Test

    >> Barcode Management

     

    Dear customer, if you send your samples via our dropboxes, please make sure to order the orange stickers below directly with your barcodes. This will help to reduce our carbon footprint. Thank you very much.

     

     


    Prepaid MPC Barcode

    Mycoplasmacheck Barcodes

    • Order your barcodes in advance
    • Label your samples and benefit from secure sample assignment
    • No order needs to be placed when sending the samples
     
     
     
    Quantity:

     
     
    Price:
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    Mycoplasmacheck barcodes remain valid for five years from the date of ordering, after which they expire.

     

    Want to use our Dropboxes to send your samples?

    Please order the stickers below and put them on the bag / box where your samples are shipped in  (within the envelope). This will ensure that your samples are routed savely and fast to our mycoplasmacheck testing laboratory.

    Please find your closest dropbox location here >>

     


    Stickers for your Tube Bags

    Sticker to send your MPC samples via our Dropboxes


    Convenient sample shipment.

    • One unit includes 10 shipment stickers
    • You only need one sticker per shipment
     
     
     
    Quantity:

     
     
    Price:
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    Why should I test my cell lines for Mycoplasma contamination?

     

    As contamination of mycoplasma alters cell-line properties, their unnoticed presence often leads to non-reproducible results. Regular testing with Mycoplasmacheck serves as a rapid, accurate and objective means of checking cells for mycoplasma contamination, ensuring cell lines are kept reliably clean.

    Highlights of the Mycoplasmacheck Service:

     

    • Standardised qPCR test for mycoplasma
    • Data provided online within 48 hours after sample receipt
    • Test history report for select periods of time can be compiled online at any time

     

    >> FAQ's MPC

    >> MPC Results

     

     

     Product details & sample submission

     

     

    Product Details

    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

     

     

    Additional Information

    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

     

     

    Publications

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

     

    2024

    Mycoplasmacheck: Pierre Joanne, Yeranuhi Hovhannisyan, Alexandre Simon, Gaëlle Revet, Romain Diot, Gabriel Friob, Denisa Calin, Zhenlin Li, Anthony Béhin, Karim Wahbi, Gérard Tachdjian, Onnik Agbulut, Generation of human induced pluripotent stem cell lines from five patients with Myofibrillar myopathy carrying different heterozygous mutations in the DES gene, Stem Cell Research 

    Mycoplasmacheck: Marta Ribeiro, Sandra Martins, Teresa Carvalho, Marta Furtado, Joaquim Sampaio Cabral, Dulce Brito, Maria Carmo-Fonseca, Simão Teixeira da Rocha,Generation of induced pluripotent stem cell lines from two unrelated individuals with familial hypertrophic cardiomyopathy carrying the MYBPC3 missense c.1484G>A mutation, Stem Cell Research

    Mycoplasmacheck: Laetitia Duboscq-Bidot, Bénédicte Hoareau, Flavie Ader, Vincent Fontaine, Eric Villard, Generation of CRISPR-Cas9 edited human induced pluripotent stem cell line carrying BAG3 V468M mutation in its BAG domain, Stem Cell Research

    Mycoplasmacheck: Ujevic, A., Knizkova, D., Synackova, A. et al. TBK1-associated adapters TANK and AZI2 protect mice against TNF-induced cell death and severe autoinflammatory diseases. Nat Commun

    Mycoplasmacheck: Jacquemin, C., El Orch, W., Diaz, O. et al. Pharmacological induction of the hypoxia response pathway in Huh7 hepatoma cells limits proliferation but increases resilience under metabolic stress. Cell. Mol. Life Sci.

    Mycoplasmacheck: Derouane, F., Desgres, M., Moroni, C. et al. Metabolic adaptation towards glycolysis supports resistance to neoadjuvant chemotherapy in early triple negative breast cancers. Breast Cancer Res

     

    2023

    Mycoplasmacheck: Abouzayed A, Zedan W, Altai M, Strand J, Örbom A. Co-injection of anti-HER2 antibody Trastuzumab does not increase efficacy of [177Lu]Lu-PSMA-617 therapy in an animal model of prostate cancer. Am J Nucl Med Mol Imaging.

    Mycoplasmacheck: Marianne Burbage et al. ,Epigenetically controlled tumor antigens derived from splice junctions between exons and transposable elements.Sci. Immunol.

    Mycoplasmacheck: Sarah Ceylan Krassnig, Marina Mäser, Nicola Anna Probst, Jens Werner, Charlotte Schlett, Nina Schumann, Gudrun von Scheven, Aswin Mangerich, Alexander Bürkle, Comparative analysis of chlorambucil-induced DNA lesion formation and repair in a spectrum of different human cell systems, Toxicology Reports

    Mycoplasmacheck: Pandiloski, N., Horváth, V., Karlsson, O. et al. DNA methylation governs the sensitivity of repeats to restriction by the HUSH-MORC2 corepressor. Nat Commun

     

    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

     

     

     

    Sample preparation

    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

     

    Sample preparation of cell cultures:

     

    • Transfer 500 μl of cell culture supernatant from the test cell culture to a 1.5 ml tube. The lid should be sealed tightly to prevent opening during heating.
    • For suspension cell lines, stand culture flasks vertically allowing cells to settle for about 30 minutes prior to removal of 500μL of the supernatant and transfer to a 1.5 ml tube.
    • Boil supernatant at 95 °C for 10 minutes.
    • Briefly centrifuge (5 seconds) the sample at ~13,000 rpm to pellet cellular debris.
    • Transfer 100 – 200 μl of supernatant into a new 1.5 ml tube labelled with a MYCOPLASMACHECK barcode. Do not disturb pellet

    Sample preparation of cell cultures that are prone to PCR inhibition:

     

    For adherent cell lines:

     

    • Transfer 500 μl of cell culture supernatant from the cell culture to a 1.5 ml tube.
    • Centrifuge sample for 10 min at >10.000 x g to pellet mycoplasma cells.
    • Discard 450 μL of supernatant without disturbing any pellet.
    • Boil remaining 50μL at 95 °C for 10 minutes. The lid should be sealed tightly to prevent
      opening during heating.
    • Perform DNA extraction using any commercially available DNA extraction kits according to
      the manufacturer ́s instructions.
    • Elute in 30 μL using a 1.5 ml tube labelled with a MYCOPLASMACHECK barcode.

     

    For suspension cell lines:

     

    • Centrifuge 500 μL sample for 3 min at >300 x g to pellet eucaryotic cells.
    • Transfer supernatant in new 1.5 ml tube.
    • Continue with 2nd step above.

     

     

    Sending samples

    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 Food/Environment/White Biotech Products & Services GmbH
    FEB species determination
    Anzinger Strasse 7a
    85560 Ebersberg

     


    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.

     

    1

    Only use 1.5 ml
    safe-lock tubes

     

    2

    Remove barcode
    stricker from film

     

    3

    Affix barcode sticker
    horizontally

     

    4

    QR-code visible at front so
    it can be read by scaner

    Order Management

    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

     

     

     Literature

     

    Mycoplasmacheck sample submission guide

    Application note

    Mycoplasma species detected

     

     

     

     

     

     

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