An assessment of air as a source of DNA contamination encountered when performing PCR.
Sensitive molecular methods, such as the PCR, can detect low-level contamination, and careful technique is required to reduce the impact of contaminants. Yet, some assays that are designed to detect high copy-number target sequences appear to be impossible to perform without contamination, and frequently, personnel or laboratory environment are held responsible as the source. This complicates diagnostic and research analysis when using molecular methods. To investigate the air specifically as a source of contamination, which might occur during PCR setup, we exposed tubes of water to the air of a laboratory and clean hood for up to 24 h. To increase the chances of contamination, we also investigated a busy open-plan office in the same way. All of the experiments showed the presence of human and rodent DNA contamination. However, there was no accumulation of the contamination in any of the environments investigated, suggesting that the air was not the source of contamination. Even the air from a busy open-plan office was a poor source of contamination for all of the DNA sequences investigated (human, bacterial, fungal, and rodent). This demonstrates that the personnel and immediate laboratory environment are not necessarily to blame for the observed contamination.
Witt
Nina
N
Centre for Infectious Diseases and International Health, Windeyer Institute for Medical Sciences, University College London, London, United Kingdom.
Rodger
Gillian
G
Vandesompele
Jo
J
Benes
Vladimir
V
Zumla
Alimuddin
A
Rook
Graham A
GA
Huggett
Jim F
JF
D000388:Air; D000392:Air Movements; D000393:Air Pollutants; D020087:Alu Elements; D000818:Animals; D001483:Base Sequence; D002175:Candida / Q000378:metabolism; D004247:DNA / Q000032:analysis*; D004275:DNA, Ribosomal / Q000235:genetics; D004784:Environmental Monitoring / Q000379:methods; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012689:Sequence Homology, Nucleic Acid
D000393:Air Pollutants; D004275:DNA, Ribosomal; D004247:DNA