关键词: 聚合酶链式反应, 细胞培养, 种属鉴定

Abstract: Objective To identify the species of cultured cells and to detect inter-species cross contamination by polymerase chain reaction (PCR). Methods From references and NCBI database, we commercially synthesized 32 pairs of species-specific primers targeted to genomic DNA of ten common species of cultured cells. Then we screened for optional primers with good specificity and high sensitivity. PCR amplification with these primers and the genomic DNAs isolated from the tested cell line, and agarose gel electrophoresis of the PCR products were done. Mixed DNA of 10 species was used as positive-control template, and water as negative-control template. Results Finally 10 pairs of species-specific and high sensitive primers were selected. By PCR amplification with these primers and agarose gel electrophoresis, we could easily tell the origin of cell lines and detect whether the tested cell lines are contaminated by cells of other species. Conclusions This PCR assay provides a simple, rapid, sensitive, and cost-effective method to identify cell species and detect interspecies cross-contamination.

Key words: polymerase chain reaction, cell culture, species identification