title: A simple preparation of mycoplasmal DNA template for PCR from biological samples using effective surfactants.
abstract: To prepare mycoplasmal DNA template for PCR from biological samples rapidly and easily, surfactants which can solubilize cell membranes effectively were investigated. 3-[3-Cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS) was considered an effective surfactant. This surfactant could solubilize mycoplasma cell membrane without suppressing the polymerase reaction. In addition, proteinase K treatment played an important role in preparing mycoplasmal DNA template from a simulated biological sample. It was therefore considered that a combination of proteinase K- and CHAPS- added lysis buffer would be more useful in preparing mycoplasmal DNA template. We could detect PCR products by using the lysis buffer with a simulated lung emulsion sample containing mycoplasma organisms at 10(4) CFU per gram.
authors: National Institute of Animal Health, Ibaraki, Japan.
authors: H
authors: H
authors: Kobayashi
authors: G
authors: G
authors: Munthali
authors: C
authors: C
authors: Miyamoto
authors: T
authors: T
authors: Morozumi
authors: K
authors: K
authors: Mitani
authors: N
authors: N
authors: Ito
authors: H
authors: H
authors: Shiono
authors: K
authors: K
authors: Yamamoto
mesh_terms: D000818:Animals; D002793:Cholic Acids; D017931:DNA Primers; D004269:DNA, Bacterial / Q000302:isolation & purification*; D005227:Fatty Acids; D005944:Glucosamine / Q000031:analogs & derivatives; D009174:Mycoplasma / Q000235:genetics / Q000302:isolation & purification*; D009175:Mycoplasma Infections / Q000175:diagnosis / Q000662:veterinary*; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000662:veterinary; D013501:Surface-Active Agents; D013552:Swine; D013553:Swine Diseases; D013698:Templates, Genetic
chemical_list: D002793:Cholic Acids; D017931:DNA Primers; D004269:DNA, Bacterial; D005227:Fatty Acids; D013501:Surface-Active Agents; C048531:chapso; C042221:decanoyl-N-methylglucamide; D005944:Glucosamine; C028213:3-((3-cholamidopropyl)dimethylammonium)-1-propanesulfonate