title: Detection of Mycoplasma Contamination Directly from Culture Supernatant Using Polymerase Chain Reaction.
abstract: Ensuring mycoplasma-free cell culture is of prime importance as they severely affect cellular characteristics leading to experimental artefacts and spurious results. Various methods persist for mycoplasma detection; out of the whole array of methods polymerase chain reaction (PCR) is the most favoured one because it is highly sensitive, specific and quick. The PCR-based detection procedure involves three steps: cell culture supernatant collection, DNA isolation, and PCR. We have modified this procedure so that cell culture supernatant can directly be used for PCR without the need for DNA extraction. This modification makes the procedure quicker and more sensitive because loss of mycoplasma DNA is prevented and this loss becomes more significant when the level of mycoplasma contamination is very low.
authors.affiliation: Department of Histology and Embryology, Charles University - Faculty of Medicine in Hradec Králové.
authors.forename: R V
authors.initials: RV
authors.lastname: Pisal
authors.affiliation: Department of Histology and Embryology, Charles University - Faculty of Medicine in Hradec Králové.
authors.forename: H
authors.initials: H
authors.lastname: Hrebíková
authors.affiliation: Department of Histology and Embryology, Charles University - Faculty of Medicine in Hradec Králové.
authors.forename: J
authors.initials: J
authors.lastname: Chvátalová
authors.affiliation: Department of Histology and Embryology, Charles University - Faculty of Medicine in Hradec Králové.
authors.forename: D
authors.initials: D
authors.lastname: Kunke
authors.affiliation: Department of Oncology and Radiotherapy, Charles University - Faculty of Medicine in Hradec Králové.
authors.forename: S
authors.initials: S
authors.lastname: Filip
authors.affiliation: Department of Histology and Embryology, Charles University - Faculty of Medicine in Hradec Králové.
authors.forename: J
authors.initials: J
authors.lastname: Mokrý
mesh_terms: D000818:Animals; D002460:Cell Line; D059018:DNA Contamination; D006801:Humans; D051379:Mice; D009174:Mycoplasma / Q000235:genetics* / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*