Allelic dropout from a high-quality DNA source |
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Authors: | Carl D Soulsbury Graziella Iossa Keith J Edwards Philip J Baker Stephen Harris |
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Institution: | (1) School of Biological Sciences, University of Bristol, Woodland Road, Bristol, BS8 1UG, UK |
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Abstract: | Allelic dropouts are an important source of genotyping error, particularly in studies using non-invasive sampling techniques.
This has important implications for conservation biology, as an increasing number of studies are now using non-invasive techniques
to study rare species or endangered populations. Previously, allelic dropout has typically been associated with PCR amplification
of low quality/quantity template DNA. However, in this study we recorded high levels of allelic dropout (21–57%) at specific
loci amplified from a high quality DNA (63.1 ± 7.8 ng/μl) source in the red fox (Vulpes vulpes). We designed a series of experiments to identify the sources of error. Whilst we were able to show that the best method
to identify allelic dropout was the dilution of template DNA prior to PCR amplification, our data also showed two specific
patterns: (1) allelic dropouts occurred at specific loci; (2) allelic dropouts occurred at specific pair-wise combinations
of alleles. These patterns suggest that mechanisms other than low quantity template DNA are responsible for allelic dropout.
Further research on the causes of these patterns in this and other studies would further our understanding of genotyping errors
and would aid future studies where allelic dropout may be a serious issue. |
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Keywords: | Microsatellites Cross-species Parentage Demography Red fox Vulpes |
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