A method to identify a large number of mammalian species in the UK from trace samples and mixtures without the use of sequencing

Shanan S. Tobe, Adrian Linacre

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

There is no standard test to identify the species of origin of a sample. A general method is to amplify part of the mitochondrial genome, generally the 12S, 16S or cytochrome b gene, and sequence it for comparison with known sequences on GenBank. Highly degraded samples and mixtures make this technique unsuitable.

As a functioning protein, cytochrome b cannot mutate unconditionally. Detrimental changes in the amino acid sequence or composition will result in cell death and would not be passed on to offspring. By examining the cytochrome b sequences non-detrimental variation can be found which can be used for specific-species identification. Areas of high homology can also be identified for universal amplification sites.

Species-specific primers have been developed based on these SNPs in the cytochrome b gene such that they will only react for a particular species. By combining universal priming sites with species-specific sites, a simple yet effect test has been constructed for the identification of species. This test will produce a product of a particular size for each species. It will work on mixtures and has sensitivity to the femtogramme (10−15 g) level.

Fingerprint

Cytochromes b
Mitochondrial Genome
Nucleic Acid Databases
Genes
Single Nucleotide Polymorphism
Amino Acid Sequence
Cell Death
Proteins

Keywords

  • mammalian mtDNA
  • non-human DNA
  • trace evidence
  • cytochrome b

Cite this

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title = "A method to identify a large number of mammalian species in the UK from trace samples and mixtures without the use of sequencing",
abstract = "There is no standard test to identify the species of origin of a sample. A general method is to amplify part of the mitochondrial genome, generally the 12S, 16S or cytochrome b gene, and sequence it for comparison with known sequences on GenBank. Highly degraded samples and mixtures make this technique unsuitable. As a functioning protein, cytochrome b cannot mutate unconditionally. Detrimental changes in the amino acid sequence or composition will result in cell death and would not be passed on to offspring. By examining the cytochrome b sequences non-detrimental variation can be found which can be used for specific-species identification. Areas of high homology can also be identified for universal amplification sites. Species-specific primers have been developed based on these SNPs in the cytochrome b gene such that they will only react for a particular species. By combining universal priming sites with species-specific sites, a simple yet effect test has been constructed for the identification of species. This test will produce a product of a particular size for each species. It will work on mixtures and has sensitivity to the femtogramme (10−15 g) level.",
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A method to identify a large number of mammalian species in the UK from trace samples and mixtures without the use of sequencing. / Tobe, Shanan S.; Linacre, Adrian.

In: Forensic Science International: Genetics Supplement Series, Vol. 1, No. 1, 08.2008, p. 625-627.

Research output: Contribution to journalArticle

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AU - Linacre, Adrian

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