Open AccessDecomposition of waste DNA with extended autoclaving under unsaturated steam
Author(s) -
Tetsushi Suyama,
Mamoru Kawaharasaki
Publication year - 2013
Publication title -
biotechniques/biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/000114113
Subject(s) - autoclave , contamination , dna , sterilization (economics) , pulp and paper industry , decomposition , waste management , environmental science , chemistry , biology , chromatography , environmental chemistry , biochemistry , organic chemistry , ecology , monetary economics , economics , foreign exchange market , engineering , foreign exchange
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.