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Residual DNA in thermostable DNA polymerases – a cause of irritation in diagnostic PCR and microarray assays
Ehricht, Ralf ; Hotzel, Helmut ; Sachse, Konrad ; Slickers, Peter
Biologicals, 2007-04, Vol.35 (2), p.145-147
[Peer Reviewed Journal]
England: Elsevier Ltd
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Title:
Residual DNA in thermostable DNA polymerases – a cause of irritation in diagnostic PCR and microarray assays
Author:
Ehricht, Ralf
;
Hotzel, Helmut
;
Sachse, Konrad
;
Slickers, Peter
Subjects:
Chlamydiaceae - genetics
;
Chlamydiaceae - isolation & purification
;
Diagnostic microarray
;
DNA
;
DNA, Bacterial - analysis
;
DNA, Bacterial - isolation & purification
;
DNA-Directed DNA Polymerase - chemistry
;
DNA-Directed DNA Polymerase - metabolism
;
Equipment Contamination
;
Escherichia coli
;
Escherichia coli - genetics
;
Escherichia coli - isolation & purification
;
Microarray Analysis - standards
;
PCR
;
Polymerase
;
Polymerase Chain Reaction - standards
;
Reagent Kits, Diagnostic - microbiology
;
Sensitivity and Specificity
;
Taq Polymerase - chemistry
;
Temperature
Is Part Of:
Biologicals, 2007-04, Vol.35 (2), p.145-147
Notes:
ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-2
content type line 23
Description:
In a validation trial of a DNA microarray test for chlamydiae we repeatedly observed false-positive PCR amplicons from truly negative samples and non-template controls. Various PCR tests, microarray hybridization and DNA sequencing, revealed that residual Escherichia coli DNA from thermostable DNA polymerases was the cause of this cross-reaction. A subsequent survey showed that only five out of 23 commercial polymerases were free of E. coli DNA. When designing generic oligonucleotide sequences for PCR and PCR microarray-based assays one should be aware of such possible internal contamination, particularly when the target organism is E. coli.
Publisher:
England: Elsevier Ltd
Language:
English
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