title: Decomposition of waste DNA with extended autoclaving under unsaturated steam.
abstract: Carryover and false-positive amplification of undesired DNA sequences are serious problems in research and diagnostic testing using PCR. One possible source of DNA cross-contamination can be the autoclave if DNA contained in waste is not effectively decomposed and contaminates the autoclave. To assess this possibility, we used a 2682 bp PCR product as a model waste DNA and quantified the amplifiability of an 84 bp short fragment derived from the model waste DNA in the steam and the residual bottom water after autoclaving. Autoclaving under the standard conditions of 121°C for 20 min did not sufficiently remove amplifiability from the model DNA and was found to be a possible source of laboratory contamination. However, the amplifiable template was removed after autoclaving at 121°C for 80 min. Fragmentation and hydrolysis may occur during autoclaving, and the presence of atmospheric oxygen facilitated the decomposition. These findings will help researchers develop better strategies for disposing of DNA waste.
authors.affiliation: National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Ibaraki, Japan.
authors.forename: Tetsushi
authors.initials: T
authors.lastname: Suyama
authors.affiliation: National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Ibaraki, Japan.
authors.forename: Mamoru
authors.initials: M
authors.lastname: Kawaharasaki
mesh_terms: D004247:DNA / Q000737:chemistry*; D006358:Hot Temperature; D016133:Polymerase Chain Reaction; D013227:Steam; D013242:Sterilization / Q000379:methods*
chemical_list: D013227:Steam; D004247:DNA
keywords: DNA waste; PCR; autoclave; contamination; decomposition; false-positive