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Synergy between DNA polymerases increases polymerase chain reaction inhibitor tolerance in forensic DNA analysis.

Hedman, Johannes LU ; Nordgaard, Anders; Dufva, Charlotte; Rasmusson, Birgitta; Ansell, Ricky and Rådström, Peter LU (2010) In Analytical Biochemistry 405(2). p.192-200
Abstract
The success rate of diagnostic polymerase chain reaction (PCR) analysis is lowered by inhibitory substances present in the samples. Recently, we showed that tolerance to PCR inhibitors in crime scene saliva stains can be improved by replacing the standard DNA polymerase AmpliTaq Gold with alternative DNA polymerase-buffer systems (Hedman et al., BioTechniques 47 (2009) 951-958). Here we show that blending inhibitor-resistant DNA polymerase-buffer systems further increases the success rate of PCR for various types of real crime scene samples showing inhibition. For 34 of 42 "inhibited" crime scene stains, the DNA profile quality was significantly improved using a DNA polymerase blend of ExTaq Hot Start and PicoMaxx High Fidelity compared... (More)
The success rate of diagnostic polymerase chain reaction (PCR) analysis is lowered by inhibitory substances present in the samples. Recently, we showed that tolerance to PCR inhibitors in crime scene saliva stains can be improved by replacing the standard DNA polymerase AmpliTaq Gold with alternative DNA polymerase-buffer systems (Hedman et al., BioTechniques 47 (2009) 951-958). Here we show that blending inhibitor-resistant DNA polymerase-buffer systems further increases the success rate of PCR for various types of real crime scene samples showing inhibition. For 34 of 42 "inhibited" crime scene stains, the DNA profile quality was significantly improved using a DNA polymerase blend of ExTaq Hot Start and PicoMaxx High Fidelity compared with AmpliTaq Gold. The significance of the results was confirmed by analysis of variance. The blend performed as well as, or better than, the alternative DNA polymerases used separately for all tested sample types. When used separately, the performance of the DNA polymerases varied depending on the nature of the sample. The superiority of the blend is discussed in terms of complementary effects and synergy between the DNA polymerase-buffer systems. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Analytical Biochemistry
volume
405
issue
2
pages
192 - 200
publisher
Elsevier
external identifiers
  • wos:000281296300007
  • pmid:20599651
  • scopus:77955564503
ISSN
1096-0309
DOI
10.1016/j.ab.2010.06.028
language
English
LU publication?
yes
id
2deda228-928e-4a22-bf6e-14dc2c0152d3 (old id 1645300)
date added to LUP
2010-08-18 12:39:08
date last changed
2018-05-29 12:20:02
@article{2deda228-928e-4a22-bf6e-14dc2c0152d3,
  abstract     = {The success rate of diagnostic polymerase chain reaction (PCR) analysis is lowered by inhibitory substances present in the samples. Recently, we showed that tolerance to PCR inhibitors in crime scene saliva stains can be improved by replacing the standard DNA polymerase AmpliTaq Gold with alternative DNA polymerase-buffer systems (Hedman et al., BioTechniques 47 (2009) 951-958). Here we show that blending inhibitor-resistant DNA polymerase-buffer systems further increases the success rate of PCR for various types of real crime scene samples showing inhibition. For 34 of 42 "inhibited" crime scene stains, the DNA profile quality was significantly improved using a DNA polymerase blend of ExTaq Hot Start and PicoMaxx High Fidelity compared with AmpliTaq Gold. The significance of the results was confirmed by analysis of variance. The blend performed as well as, or better than, the alternative DNA polymerases used separately for all tested sample types. When used separately, the performance of the DNA polymerases varied depending on the nature of the sample. The superiority of the blend is discussed in terms of complementary effects and synergy between the DNA polymerase-buffer systems.},
  author       = {Hedman, Johannes and Nordgaard, Anders and Dufva, Charlotte and Rasmusson, Birgitta and Ansell, Ricky and Rådström, Peter},
  issn         = {1096-0309},
  language     = {eng},
  number       = {2},
  pages        = {192--200},
  publisher    = {Elsevier},
  series       = {Analytical Biochemistry},
  title        = {Synergy between DNA polymerases increases polymerase chain reaction inhibitor tolerance in forensic DNA analysis.},
  url          = {http://dx.doi.org/10.1016/j.ab.2010.06.028},
  volume       = {405},
  year         = {2010},
}