Literature DB >> 2087233

Taq polymerase contains bacterial DNA of unknown origin.

K H Rand1, H Houck.   

Abstract

The polymerase chain reaction (PCR) was carried out with the highly conserved E. coli ribosomal RNA gene sequences 1376-1395 and 1521-1540. Using these primers and reaction conditions specified by the manufacturer(s), a 165 bp fragment was synthesized using Taq polymerase from three different sources in the absence of any added template. Restriction enzyme analysis suggests the source of this bacterial DNA is neither E. coli nor Thermus aquaticus. A variety of different methods to eliminate it such as treatment with DNase, restriction enzyme digestion, and CsCl2 density gradient centrifugation were unsuccessful. Since the bacteria in which the Taq polymerase is produced are not the source of the DNA, some step(s) in the purification or reagents added to the enzyme must be involved. Thus it is likely other biological products are similarly contaminated. Although the problem is easily dealt with by running a no-template control and choosing other primers if a problem exists, it is important to recognize the potential for a false-positive result.

Entities:  

Mesh:

Substances:

Year:  1990        PMID: 2087233     DOI: 10.1016/0890-8508(90)90003-i

Source DB:  PubMed          Journal:  Mol Cell Probes        ISSN: 0890-8508            Impact factor:   2.365


  45 in total

1.  Does blood of healthy subjects contain bacterial ribosomal DNA?

Authors:  S Nikkari; I J McLaughlin; W Bi; D E Dodge; D A Relman
Journal:  J Clin Microbiol       Date:  2001-05       Impact factor: 5.948

2.  Contamination and sensitivity issues with a real-time universal 16S rRNA PCR.

Authors:  C E Corless; M Guiver; R Borrow; V Edwards-Jones; E B Kaczmarski; A J Fox
Journal:  J Clin Microbiol       Date:  2000-05       Impact factor: 5.948

3.  DNase pretreatment of master mix reagents improves the validity of universal 16S rRNA gene PCR results.

Authors:  Alexandra Heininger; Marlies Binder; Andreas Ellinger; Konrad Botzenhart; Klaus Unertl; Gerd Döring
Journal:  J Clin Microbiol       Date:  2003-04       Impact factor: 5.948

4.  Real-time PCR for detection and differentiation of gram-positive and gram-negative bacteria.

Authors:  Sven Klaschik; Lutz E Lehmann; Ansgar Raadts; Malte Book; Andreas Hoeft; Frank Stuber
Journal:  J Clin Microbiol       Date:  2002-11       Impact factor: 5.948

Review 5.  Next-generation sequencing in the analysis of human microbiota: essential considerations for clinical application.

Authors:  Geraint B Rogers; Kenneth D Bruce
Journal:  Mol Diagn Ther       Date:  2010-12-01       Impact factor: 4.074

6.  Prospective study of use of PCR amplification and sequencing of 16S ribosomal DNA from cerebrospinal fluid for diagnosis of bacterial meningitis in a clinical setting.

Authors:  Tim Schuurman; Richard F de Boer; Anna M D Kooistra-Smid; Anton A van Zwet
Journal:  J Clin Microbiol       Date:  2004-02       Impact factor: 5.948

7.  Comparison of different decontamination methods for reagents to detect low concentrations of bacterial 16S DNA by real-time-PCR.

Authors:  Sven Klaschik; Lutz E Lehmann; Ansgar Raadts; Andreas Hoeft; Frank Stuber
Journal:  Mol Biotechnol       Date:  2002-11       Impact factor: 2.695

8.  Presence of bacterial phage-like DNA sequences in commercial Taq DNA polymerase reagents.

Authors:  Tamara Newsome; Bing-Jie Li; Nianxiang Zou; Shyh-Ching Lo
Journal:  J Clin Microbiol       Date:  2004-05       Impact factor: 5.948

Review 9.  False-positive results and contamination in nucleic acid amplification assays: suggestions for a prevent and destroy strategy.

Authors:  A Borst; A T A Box; A C Fluit
Journal:  Eur J Clin Microbiol Infect Dis       Date:  2004-03-10       Impact factor: 3.267

10.  Obstacles of Multiplex Real-Time PCR for Bacterial 16S rDNA: Primer Specifity and DNA Decontamination of Taq Polymerase.

Authors:  Sebastian Philipp; Hartwig P Huemer; Eveline U Irschick; Christoph Gassner
Journal:  Transfus Med Hemother       Date:  2010-01-07       Impact factor: 3.747

View more

北京卡尤迪生物科技股份有限公司 © 2022-2023.