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Introduction

Contaminations of cell culture pose a major threat to scientific results. In addition to molds, yeast and bacteria contaminations that are easily detectable by pH shifts, turbidity, and cell destruction, other contaminations such as viruses, mycoplasma and other cell lines are more difficult to detect, and as a result, are a potentially more serious problem.

The DKFZ has developed a high-throughput multiplex cell contamination test (McCT) currently able to detect >20 contamination markers in a single reaction.

Non-DKFZ members have access to this McCT service through the Steinbeis-Transfer Centre Multiplexion.

 

Multiplex cell Contamination Testing (McCT) v.2 Service

The McCT v.2 service is designed to simultaneously detect contaminations with 

 

What is new in McCT v.2?

The latest version of our service also tests for Syrian hamster and Drosophila cell lines. None-exclusive cell line markers including HPV18, EBV and others, are not further tested for cell line authentication. The Multiplex human Cell line Authentication service (MCA) is available here.

 

Test specifications

Detection of contaminations after DNA extraction is done with specific primer sequences in a multiplex PCR targeting viral, bacterial and cellular genome regions, respectively, and subsequent hybridisation using specific oligonucleotide probes. For internal control of DNA quality the presence of the PolA gene is tested as a reference. Several positive and negative controls are included to test PCR performance.

 

Contamination Control Procedure

To be done by the user

  • Cell lysis of cell line(s) by boiling (protocol)
  • Complete the Contamination Control Submission Form (Login required)
  • Label tubes with 'KstSt' AND 'specific sample name' as provided in the Submission Form
  • Delivery of lysate(s) to the ATV building INF 242
    every Monday*, 10-12 a.m. to the doorman

* Attention: The McCT service will be performed on the first Monday each month! Lysates can be delivered weekly every Monday (no delivery between December 27 to January 7). If Monday is a bank holiday, lysates should be delivered on Tuesdays. In January 2012 no testing will be performed. Starting from January 2012, new prices are applicable (price list, login required).

  

To be done by the Service Unit

  • Multiplex PCR-based amplification of contaminants
  • Luminex-based detection of PCR products by specific oligonucleotide probes
  • Transfer of results to customer

  

Mycoplasma elimination

We recommend to follow our protocol to eliminate Mycoplasma from your precious cell lines.

 

Recommended frequency of testing

According to our own experience, we recommend to control

  • All batches of stocked cell lines once
  • All new cell lines and new virus cultures
  • Long term-cultured cell lines every month
  • In case of mycoplasma therapy every week
  • Prior to or concomitant to storage in liquid nitrogen
  • In case of alterations in cell features
  • In case of problems with reproducibility of results

 

Reference: Schmitt, M., and M. Pawlita. 2009. High-throughput detection and multiplex identification of cell contaminations. Nucleic Acids Res 37:e119. [PDF].

When citing this service, please use the above mentioned reference and indicate the time that passed between the experiments described in your manuscript and the McCT service.

 

Please note: This DKFZ-internal service is designed to help you comply with safety regulations, to improve quality control and to help your research. It is NOT suited for clinical diagnostics.

For more information, please contact us.

For detailed cost information please consult the price list (login required).

last update:
12/12/2011
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