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Improved forensic DNA analysis through the use of alternative DNA polymerases and statistical modeling of DNA profiles.

Hedman, Johannes LU ; Nordgaard, Anders; Rasmusson, Birgitta; Ansell, Ricky and Rådström, Peter LU (2009) In BioTechniques 47(5). p.951-958
Abstract
DNA evidence, linking perpetrators to crime scenes, is central to many legal proceedings. However, DNA samples from crime scenes often contain PCR-inhibitory substances, which may generate blank or incomplete DNA profiles. Extensive DNA purification can be required to rid the sample of these inhibitors, although these procedures increase the risk of DNA loss. Most forensic laboratories use commercial DNA amplification kits (e.g., AmpFlSTR SGM Plus) with the DNA polymerase AmpliTaq Gold as the gold standard. Here, we show that alternative DNA polymerase-buffer systems can improve the quality of forensic DNA analysis and efficiently circumvent PCR inhibition in crime scene samples, without additional sample preparation. DNA profiles from 20... (More)
DNA evidence, linking perpetrators to crime scenes, is central to many legal proceedings. However, DNA samples from crime scenes often contain PCR-inhibitory substances, which may generate blank or incomplete DNA profiles. Extensive DNA purification can be required to rid the sample of these inhibitors, although these procedures increase the risk of DNA loss. Most forensic laboratories use commercial DNA amplification kits (e.g., AmpFlSTR SGM Plus) with the DNA polymerase AmpliTaq Gold as the gold standard. Here, we show that alternative DNA polymerase-buffer systems can improve the quality of forensic DNA analysis and efficiently circumvent PCR inhibition in crime scene samples, without additional sample preparation. DNA profiles from 20 of 32 totally or partially inhibited crime scene saliva samples were significantly improved using Bio-X-Act Short, ExTaq Hot Start, or PicoMaxx High Fidelity instead of AmpliTaq Gold. A statistical model for unbiased quality control of forensic DNA profiles was developed to quantify the results. Our study demonstrates the importance of adjusting the chemistry of the PCR to enhance forensic DNA analysis and diagnostic PCR, providing an alternative to laborious sample preparation protocols. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
BioTechniques
volume
47
issue
5
pages
951 - 958
publisher
Informa Healthcare
external identifiers
  • wos:000277560200010
  • pmid:20041848
  • scopus:77949320758
ISSN
1940-9818
DOI
10.2144/000113246
language
English
LU publication?
yes
id
15467766-3f3b-45f9-9d3c-1fb1fef325cc (old id 1541459)
date added to LUP
2010-02-02 15:46:44
date last changed
2017-11-19 03:58:47
@article{15467766-3f3b-45f9-9d3c-1fb1fef325cc,
  abstract     = {DNA evidence, linking perpetrators to crime scenes, is central to many legal proceedings. However, DNA samples from crime scenes often contain PCR-inhibitory substances, which may generate blank or incomplete DNA profiles. Extensive DNA purification can be required to rid the sample of these inhibitors, although these procedures increase the risk of DNA loss. Most forensic laboratories use commercial DNA amplification kits (e.g., AmpFlSTR SGM Plus) with the DNA polymerase AmpliTaq Gold as the gold standard. Here, we show that alternative DNA polymerase-buffer systems can improve the quality of forensic DNA analysis and efficiently circumvent PCR inhibition in crime scene samples, without additional sample preparation. DNA profiles from 20 of 32 totally or partially inhibited crime scene saliva samples were significantly improved using Bio-X-Act Short, ExTaq Hot Start, or PicoMaxx High Fidelity instead of AmpliTaq Gold. A statistical model for unbiased quality control of forensic DNA profiles was developed to quantify the results. Our study demonstrates the importance of adjusting the chemistry of the PCR to enhance forensic DNA analysis and diagnostic PCR, providing an alternative to laborious sample preparation protocols.},
  author       = {Hedman, Johannes and Nordgaard, Anders and Rasmusson, Birgitta and Ansell, Ricky and Rådström, Peter},
  issn         = {1940-9818},
  language     = {eng},
  number       = {5},
  pages        = {951--958},
  publisher    = {Informa Healthcare},
  series       = {BioTechniques},
  title        = {Improved forensic DNA analysis through the use of alternative DNA polymerases and statistical modeling of DNA profiles.},
  url          = {http://dx.doi.org/10.2144/000113246},
  volume       = {47},
  year         = {2009},
}