Nucleic Acids Research Advance Access published online on July 9, 2009
Nucleic Acids Research, doi:10.1093/nar/gkp581
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Methods Online |
High-throughput detection and multiplex identification of cell contaminations
Research Program Infection and Cancer, German Cancer Research Center (DKFZ), Im Neuenheimer Feld 242, 69120 Heidelberg, Germany
*To whom correspondence should be addressed. Tel: +0049 (0)6221 424937; Fax: +0049 (0)6221 424932; Email: markus.schmitt{at}dkfz.de
Received April 30, 2009. Revised June 10, 2009. Accepted June 24, 2009.
Unnoticed cell culture contamination by viruses, Mycoplasma, or other cell lines is not uncommon and a threat to laboratory safety and the quality of scientific results. We developed and validated a novel high-throughput Multiplex cell Contamination Test (McCT), which is currently able to detect 37 contamination markers in a single reaction. The assay is based on multiplex PCR with target-specific primers and subsequent hybridization of amplimers to specific oligonucleotide probes. McCT proved to be highly specific, sensitive and robust, and allows to analyze more than 1000 cell lysates per week. In conclusion, the novel McCT assay is a powerful high-throughput tool in assessing cell line purity.