title: Sequence analysis of the canine mitochondrial DNA control region from shed hair samples in criminal investigations.
abstract: In recent years, evidence from domestic dogs has increasingly been analyzed by forensic DNA testing. Especially, canine hairs have proved most suitable and practical due to the high rate of hair transfer occurring between dogs and humans. Starting with the description of a contamination-free sample handling procedure, we give a detailed workflow for sequencing hypervariable segments (HVS) of the mtDNA control region from canine evidence. After the hair material is lysed and the DNA extracted by Phenol/Chloroform, the amplification and sequencing strategy comprises the HVS I and II of the canine control region and is optimized for DNA of medium-to-low quality and quantity. The sequencing procedure is based on the Sanger Big-dye deoxy-terminator method and the separation of the sequencing reaction products is performed on a conventional multicolor fluorescence detection capillary electrophoresis platform. Finally, software-aided base calling and sequence interpretation are addressed exemplarily.
authors.affiliation: Institute of Legal Medicine, Innsbruck Medical University, Innsbruck, Austria.
authors.forename: C
authors.initials: C
authors.lastname: Berger
authors.forename: B
authors.initials: B
authors.lastname: Berger
authors.forename: W
authors.initials: W
authors.lastname: Parson
mesh_terms: D000818:Animals; D001483:Base Sequence; D002725:Chloroform; D003415:Crime; D004272:DNA, Mitochondrial / Q000235:genetics* / Q000302:isolation & purification; D004285:Dogs / Q000235:genetics*; D019075:Electrophoresis, Capillary; D053803:Forensic Genetics / Q000379:methods*; D006197:Hair / Q000378:metabolism*; D006801:Humans; D019901:Locus Control Region / Q000235:genetics*; D018598:Minisatellite Repeats / Q000235:genetics; D008969:Molecular Sequence Data; D019800:Phenol; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA / Q000379:methods*; D013223:Statistics as Topic
chemical_list: D004272:DNA, Mitochondrial; D019800:Phenol; D002725:Chloroform