Identification and elimination of DNA sequences in Taq DNA polymerase

Hughes, M S; Beck, L A; Skuce, R A

doi:10.1128/jcm.32.8.2007-2008.1994

Cited by 105Molecular Biology Techniques and ApplicationsEnzyme Production and CharacterizationGenomics and Phylogenetic Studies

Identification and elimination of DNA sequences in Taq DNA polymerase

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Hughes, M S, Beck, L A, Skuce, R A·1994·Journal of Clinical Microbiology

DOI 10.1128/jcm.32.8.2007-2008.1994PMC PMC263920

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Abstract

This study confirms that different preparations of Taq DNA polymerase are contaminated with eubacterial DNA. The contaminants appeared to represent more than one strain or species but were not identified as Thermus aquaticus or Escherichia coli. Differences in microcentrifuge tube composition appeared to affect elimination of the contaminants.

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The narrowed specificity of the FilmArray ME Panel for E. coli was a necessary design consideration to prevent detection of contaminating E. coli nucleic acid, which is common in PCR reagents.
“The narrowed specificity of the FilmArray ME Panel for this organism was a necessary design consideration in order to prevent detection of contaminating E. coli nucleic acid, which is common in PCR reagents (e.g., DNA polymerase and reverse transcriptase) (46, 47).”
From Multicenter Evaluation of BioFire FilmArray Meningitis/Encephalitis Panel for Detection of Bacteria, Viruses, and Yeast in Cerebrospinal Fluid Specimens (2016) · cites this paper as [47]

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A contamination assessment of the CI carbonaceous meteorite Orgueil using a DNA ‐directed approach
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Decontamination of polymerase chain reaction reagents using DEAE-cellulose.
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“…is repeatedly shown to be contaminated with bacterial DNA (Borst et al., 2004; Bottger, 1990; Corless et al., 2000; Goldenberger & Altwegg, 1995; Hughes et al., 1994; Lo et al., 1988; Loeffler et al., 1999; Maiwald et al., 1994; Rand & Houck, 1990; Schmidt et al., 1991). to its non-detection…”
Assessment of viable bacteria and bacterial DNA in blood and bloodstain specimens stored under various conditions.
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The Occurrence of Antibiotic Resistance Genes in Taq Polymerases and a Decontamination Method Applied to the Detection of Genetically Modified Crops
Perron, A., Raymond, P., & Simard, R. (2006) · Biotechnology Letters · doi:10.1007/s10529-005-5931-3 · Cited by 4
“Previous studies have shown that the native Taq polymerase contains DNA of unknown origin, DNA not related to Escherichia coli or Thermophilus aquaticus (Rand & Houck 1990, Hughes et al. 1994).”
Background
Application of Optical Trapping for Cells Grown on Plates: Optimization of PCR and Fidelity of DNA Sequencing of p53 Gene from a Single Cell
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“This was thought to be a result of background signal (42, 43) or PCR bias (38, 44, 45).”
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Amplification Product Inactivation
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“These methods are extremely useful if employed while doing universal 16S rRNA PCR (Hughes et al., 1994; Corless et al., 2000).”
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“The presence of contaminating DNA in commercial preparations of Taq DNA polymerase has been convin-cingly established (Bottger, 1990; Corless et al., 2000; Hughes et al., 1994; Newsome et al., 2004; Rand and Houck, 1990; Schmidt et al., 1991).”
Background
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Authors: Hughes, M. S., Beck, L. A., & Skuce, R. A.
Affiliations: Department of Agriculture for Northern Ireland, Stromont, Belfast.
ORCID: Skuce, R A
Venue: Journal of Clinical Microbiology
Publisher: American Society for Microbiology
Published: 1994-08
Volume: 32
Issue: 8
Pages: 2007-2008
ISSN: 0095-1137 (ISSN-L)
1098-660X
DOI: 10.1128/jcm.32.8.2007-2008.1994
PMC: PMC263920
OpenAlex: W1522322183
Cited by: 105
References: 0
Citations by year: 2026: 1 · 2024: 3 · 2023: 1 · 2022: 1 · 2021: 2 · 2019: 1 · 2018: 2 · 2017: 2 · 2016: 5 · 2015: 5 · 2014: 5 · 2013: 3 · 2012: 4
FWCI: 0.51
Type: journal article
Language: en
Open access: bronze
doi.org
Licence: journals.asm.org/non-commercial-tdm-license (tdm)
Locations: Journal of Clinical Microbiology (publishedVersion)
Europe PMC (PubMed Central) (publishedVersion)
PubMed Central (publishedVersion)
PubMed (publishedVersion)
Topics: Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Molecular Biology Techniques and Applications
Life Sciences › Biochemistry, Genetics and Molecular Biology › Biotechnology › Enzyme Production and Characterization
Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Phylogenetic Studies
Keywords: Taq DNA polymerase, bacterial DNA contamination, Thermus aquaticus, Escherichia coli
MeSH terms: Base Sequence, DNA-Directed DNA Polymerase, DNA, Bacterial, Drug Contamination, Methoxsalen, Molecular Sequence Data, Ultraviolet Rays, Polymerase Chain Reaction, Taq Polymerase

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Identification and elimination of DNA sequences in Taq DNA polymerase

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