†Present address: Kenneth D. Eilert, Indiana State Police Laboratory, Indianapolis, IN 46202.
Polymerase Resistance to Polymerase Chain Reaction Inhibitors in Bone*
Article first published online: 3 AUG 2009
© 2009 American Academy of Forensic Sciences
Journal of Forensic Sciences
Volume 54, Issue 5, pages 1001–1007, September 2009
How to Cite
Eilert, K. D. and Foran, D. R. (2009), Polymerase Resistance to Polymerase Chain Reaction Inhibitors in Bone. Journal of Forensic Sciences, 54: 1001–1007. doi: 10.1111/j.1556-4029.2009.01116.x
Presented at the 58th Annual Meeting of the American Academy of Forensic Sciences in San Antonio, Texas, February 19–24, 2007.
- Issue published online: 1 SEP 2009
- Article first published online: 3 AUG 2009
- Received 27 April 2008; and in revised form 19 Oct. 2008; accepted 25 Oct. 2008.
- forensic science;
- polymerase chain reaction;
- bovine serum albumin;
- Thermus aquaticus;
- Thermus ubiquitous;
- Thermus flavus;
- Thermus litoralis;
- Thermus thermophilus;
- Thermus filiformis;
- Thermococcus gorgonarius;
- Thermococcus zilligii;
- Pyrococcus woesei
Abstract: Amplification of DNA from aged or degraded skeletal remains can be a challenging task, in part due to naturally occurring inhibitors of the polymerase chain reaction. PCR inhibitors may act by inactivating a polymerase itself, or compete with or bind other reaction components, although various polymerases may be differentially susceptible to such insult. In this study, ten thermostable polymerases from six bacterial species were examined for their ability to amplify DNA in the presence of bone-derived or individual PCR inhibitors. Two polymerases, one from Thermus aquaticus and one from Thermus thermophilus, showed lower susceptibility to inhibition from bone, while polymerases from Thermus flavus were highly susceptible. Addition of bovine serum albumin improved the activity of most of the enzymes. Taken together, the results indicate that thermostable DNA polymerases have different susceptibility to bone-derived PCR inhibitors, and that those most often used in forensic laboratories may not be optimal when working with DNA from skeletal remains.