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. 2016 May;61(3):594-606.
doi: 10.1111/1556-4029.13026. Epub 2016 Jan 6.

SNP Miniplexes for Individual Identification of Random-Bred Domestic Cats

Ashley Brooks  1 , Erica K Creighton  2 , Barbara Gandolfi  1   2 , Razib Khan  1 , Robert A Grahn  1 , Leslie A Lyons  1   2
Affiliations

SNP Miniplexes for Individual Identification of Random-Bred Domestic Cats

Ashley Brooks et al. J Forensic Sci. 2016 May.

Abstract

Phenotypic and genotypic characteristics of the cat can be obtained from single nucleotide polymorphisms (SNPs) analyses of fur. This study developed miniplexes using SNPs with high discriminating power for random-bred domestic cats, focusing on individual and phenotypic identification. Seventy-eight SNPs were investigated using a multiplex PCR followed by a fluorescently labeled single base extension (SBE) technique (SNaPshot(®) ). The SNP miniplexes were evaluated for reliability, reproducibility, sensitivity, species specificity, detection limitations, and assignment accuracy. Six SNPplexes were developed containing 39 intergenic SNPs and 26 phenotypic SNPs, including a sex identification marker, ZFXY. The combined random match probability (cRMP) was 6.58 × 10(-19) across all Western cat populations and the likelihood ratio was 1.52 × 10(18) . These SNPplexes can distinguish individual cats and their phenotypic traits, which could provide insight into crime reconstructions. A SNP database of 237 cats from 13 worldwide populations is now available for forensic applications.

Keywords: Felis silvestris catus; animal forensics; forensic genetics; forensic science; single base extension; single nucleotide polymorphism.

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Figures

Figure 1
Figure 1
Electropherograms of six SNP miniplexes. Forty‐eight SNPs were successfully combined into six miniplexes. (a) represents Panel 1a, (b) represents Panel 1b, (c) represents Panel 2a, (d) represents Panel 2b, (e) represents the phenotypic Panel 3a, and (f) represents the phenotypic Panel 3b. Alleles are called as the following: A is green, T is red, G is blue, and C is black. SNP B4_40319102 did not amplify after multiplexing.
Figure 2
Figure 2
Sensitivity of the SNPs used in the six miniplexes. The SNPplexes produced complete profiles from 7.2 ng to 122.5 pg. Data for template DNA >122.5 pg not presented. Drop‐out of alleles was apparent at 56 pg and lower DNA concentrations.
Figure 3
Figure 3
Electropherograms produced by the reproducibility study (Panel 1a). (a) Represents same operator and protocol using a different capillary electrophoresis ABI 3730 DNA analyzer instrument. (b) Represents normal conditions, (c) represents different operator same instrument. Alleles are called as the following: A is green, T is red, G is blue, and C is black.
See this image and copyright information in PMC

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