Literature DB >> 19949694

An assessment of air as a source of DNA contamination encountered when performing PCR.

Nina Witt1, Gillian Rodger, Jo Vandesompele, Vladimir Benes, Alimuddin Zumla, Graham A Rook, Jim F Huggett.   

Abstract

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.

Entities:  

Keywords:  16S rDNA ; Alu repeat; B1 element; qPCR; real-time PCR

Mesh:

Substances:

Year:  2009        PMID: 19949694      PMCID: PMC2777341     

Source DB:  PubMed          Journal:  J Biomol Tech        ISSN: 1524-0215


  19 in total

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Journal:  J Forensic Sci       Date:  2000-11       Impact factor: 1.832

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Journal:  J Clin Microbiol       Date:  1999-04       Impact factor: 5.948

3.  Qiagen DNA extraction kits for sample preparation for legionella PCR are not suitable for diagnostic purposes.

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Journal:  J Clin Microbiol       Date:  2002-03       Impact factor: 5.948

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Authors:  K H Rand; H Houck
Journal:  Mol Cell Probes       Date:  1990-12       Impact factor: 2.365

5.  Whole-genome analysis of Alu repeat elements reveals complex evolutionary history.

Authors:  Alkes L Price; Eleazar Eskin; Pavel A Pevzner
Journal:  Genome Res       Date:  2004-11       Impact factor: 9.043

6.  Identification of contaminating fungal DNA sequences in Zymolyase.

Authors:  D Rimek; A P Garg; W H Haas; R Kappe
Journal:  J Clin Microbiol       Date:  1999-03       Impact factor: 5.948

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Journal:  Nature       Date:  1989-05-18       Impact factor: 49.962

8.  Use of uracil DNA glycosylase to control carry-over contamination in polymerase chain reactions.

Authors:  M C Longo; M S Berninger; J L Hartley
Journal:  Gene       Date:  1990-09-01       Impact factor: 3.688

9.  Elimination of contaminating DNA within polymerase chain reaction reagents: implications for a general approach to detection of uncultured pathogens.

Authors:  A Meier; D H Persing; M Finken; E C Böttger
Journal:  J Clin Microbiol       Date:  1993-03       Impact factor: 5.948

10.  Amplification of residual DNA sequences in sterile bronchoscopes leading to false-positive PCR results.

Authors:  K Kaul; S Luke; C McGurn; N Snowden; C Monti; W A Fry
Journal:  J Clin Microbiol       Date:  1996-08       Impact factor: 5.948

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  10 in total

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Journal:  EMBO Rep       Date:  2015-07-06       Impact factor: 8.807

2.  Progress in metagenomics requires a balanced appraisal of the available technologies.

Authors:  J F Huggett; D J Studholme; T Laver; C A Foy
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4.  Distinct DNA methylomes of newborns and centenarians.

Authors:  Holger Heyn; Ning Li; Humberto J Ferreira; Sebastian Moran; David G Pisano; Antonio Gomez; Javier Diez; Jose V Sanchez-Mut; Fernando Setien; F Javier Carmona; Annibale A Puca; Sergi Sayols; Miguel A Pujana; Jordi Serra-Musach; Isabel Iglesias-Platas; Francesc Formiga; Agustin F Fernandez; Mario F Fraga; Simon C Heath; Alfonso Valencia; Ivo G Gut; Jun Wang; Manel Esteller
Journal:  Proc Natl Acad Sci U S A       Date:  2012-06-11       Impact factor: 11.205

5.  Nasal bacterial microbiota during an outbreak of equine herpesvirus 1 at a farm in southern Ontario.

Authors:  Diego E Gomez; Luis G Arroyo; Brandon Lillie; J Scott Weese
Journal:  Can J Vet Res       Date:  2021-01       Impact factor: 1.310

6.  Proper Authentication of Ancient DNA Is Still Essential.

Authors:  Raphael Eisenhofer; Laura S Weyrich
Journal:  Genes (Basel)       Date:  2018-02-26       Impact factor: 4.096

7.  Multiplex detection of bacteria on an integrated centrifugal disk using bead-beating lysis and loop-mediated amplification.

Authors:  He Yan; Yunzeng Zhu; Yan Zhang; Lei Wang; Junge Chen; Ying Lu; Youchun Xu; Wanli Xing
Journal:  Sci Rep       Date:  2017-05-03       Impact factor: 4.379

8.  Highly sensitive and specific Alu-based quantification of human cells among rodent cells.

Authors:  Kodai Funakoshi; Mozhdeh Bagheri; Ming Zhou; Ryoji Suzuki; Hiroshi Abe; Hideo Akashi
Journal:  Sci Rep       Date:  2017-10-16       Impact factor: 4.379

Review 9.  Polymerase Chain Reaction and Its Application in the Diagnosis of Infectious Keratitis.

Authors:  Harry Y Liu; Grant C Hopping; Uma Vaidyanathan; Yasmyne C Ronquillo; Phillip C Hoopes; Majid Moshirfar
Journal:  Med Hypothesis Discov Innov Ophthalmol       Date:  2019

10.  Bacteria in the amniotic fluid without inflammation: early colonization vs. contamination.

Authors:  Eunjung Jung; Roberto Romero; Bo Hyun Yoon; Kevin R Theis; Dereje W Gudicha; Adi L Tarca; Ramiro Diaz-Primera; Andrew D Winters; Nardhy Gomez-Lopez; Lami Yeo; Chaur-Dong Hsu
Journal:  J Perinat Med       Date:  2021-07-07       Impact factor: 2.716

  10 in total

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