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

Hedman, JohannesLund University,Lunds universitet,Teknisk mikrobiologi,Centrum för tillämpade biovetenskaper,Kemiska institutionen,Institutioner vid LTH,Lunds Tekniska Högskola,Applied Microbiology,Center for Applied Life Sciences,Department of Chemistry,Departments at LTH,Faculty of Engineering, LTH,Department of Applied Microbiology, Lund University, Sweden (författare)

The success rate of diagnostic polymerase chain reaction (PCR) analysis is lowered by inhibitory substancespresent in the samples. Recently, we showed that tolerance to PCR inhibitors in crime scene salivastains can be improved by replacing the standard DNA polymerase AmpliTaq Gold with alternative DNApolymerase–buffer systems (Hedman et al., BioTechniques 47 (2009) 951–958). Here we show thatblending inhibitor-resistant DNA polymerase–buffer systems further increases the success rate of PCRfor various types of real crime scene samples showing inhibition. For 34 of 42 ‘‘inhibited” crime scenestains, the DNA profile quality was significantly improved using a DNA polymerase blend of ExTaq HotStart and PicoMaxx High Fidelity compared with AmpliTaq Gold. The significance of the results was confirmedby analysis of variance. The blend performed as well as, or better than, the alternative DNA polymerasesused separately for all tested sample types. When used separately, the performance of the DNApolymerases varied depending on the nature of the sample. The superiority of the blend is discussed interms of complementary effects and synergy between the DNA polymerase–buffer systems.

Nordgaard, AndersSwedish National Laboratory of Forencis Sciences, Linkoping, Sweden(Swepub:liu)andno60 (författare)
Dufva, CharlotteNational Laboratory of Forensic Sciences, Linkoping, Sweden (författare)
Rasmusson, BirgittaNational Laboratory of Forensic Sciences, Linkoping, Sweden(Swepub:liu)birra51 (författare)
Ansell, RickyLinköpings universitet,Molekylär genetik,Tekniska högskolan(Swepub:liu)rican78 (författare)
Rådström, PeterLund University,Lunds universitet,Teknisk mikrobiologi,Centrum för tillämpade biovetenskaper,Kemiska institutionen,Institutioner vid LTH,Lunds Tekniska Högskola,Applied Microbiology,Center for Applied Life Sciences,Department of Chemistry,Departments at LTH,Faculty of Engineering, LTH,Department of Applied Microbiology, Lund University, Sweden(Swepub:lu)tmb-pra (författare)
Teknisk mikrobiologiCentrum för tillämpade biovetenskaper (creator_code:org_t)

Ingår i:Analytical Biochemistry- : Elsevier Inc.405, s. 192-2000003-26971096-0309

Av författaren/redakt...: Hedman, Johannes; Nordgaard, Ander ...; Dufva, Charlotte; Rasmusson, Birgi ...; Ansell, Ricky; Rådström, Peter

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