title: Recovery of human DNA profiles from poached deer remains part 2: improved recovery protocol without the need for LCN analysis.
abstract: Although poaching is a common wildlife crime, the high and prohibitive cost of specialised animal testing means that many cases are left un-investigated. We previously described a novel approach to wildlife crime investigation that looked at the identification of human DNA on poached animal remains (Tobe, Govan and Welch, 2011). Human DNA was successfully isolated and amplified from simulated poaching incidents, however a low template protocol was required which made this method unsuitable for use in many laboratories. We now report on an optimised recovery and amplification protocol which removes the need for low template analysis. Samples from 10 deer (40 samples total - one from each leg) analysed in the original study were re-analysed in the current study with an additional 11 deer samples. Four samples analysed using Chelex did not show any results and a new method was devised whereby the available DNA was concentrated. By combining the DNA extracts from all tapings of the same deer remains followed by concentration, the recovered quantity of human DNA was found to be 29.5pg±43.2pg, 31× greater than the previous study. The use of the Investigator Decaplex SE (QIAGEN) STR kit provided better results in the form of more complete profiles than did the AmpFℓSTR® SGM Plus® kit at 30cycles (Applied Biosystems). Re-analysis of the samples from the initial study using the new, optimised protocol resulted in an average increase of 18% of recovered alleles. Over 17 samples, 71% of the samples analysed using the optimised protocol showed sufficient amplification for comparison to a reference profile and gave match probabilities ranging from 7.7690×10(-05) to 2.2706×10(-14). The removal of low template analysis means this optimised method provides evidence of high probative value and is suitable for immediate use in forensic laboratories. All methods and techniques used are standard and are compatible with current SOPs. As no high cost non-human DNA analysis is required the overall process is no more expensive than the investigation of other volume crime samples. The technique is suitable for immediate use in poaching incidents.
authors: Department of Pure and Applied Chemistry, University of Strathclyde, Glasgow, UK. shanan.s.tobe@strath.ac.uk
authors: Shanan S
authors: SS
authors: Tobe
authors: Stuart
authors: S
authors: Bailey
authors: James
authors: J
authors: Govan
authors: Lindsey A
authors: LA
authors: Welch
mesh_terms: D000818:Animals; D003247:Conservation of Natural Resources / Q000331:legislation & jurisprudence*; D003415:Crime / Q000331:legislation & jurisprudence*; D004247:DNA / Q000302:isolation & purification*; D016172:DNA Fingerprinting / Q000379:methods*; D003670:Deer; D006801:Humans; D018895:Microsatellite Repeats; D016133:Polymerase Chain Reaction
chemical_list: D004247:DNA