Single-tube HotSHOT technique for the collection, preservation and PCR-ready DNA preparation of faecal samples: the threatened Cabrera's vole as a model |
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Authors: | Samer Alasaad Antonio Sánchez Juan L García-Mudarra Michael J Jowers Jesús M Pérez Juan Alberto Marchal Ismael Romero José A Garrido-García Ramón C Soriguer |
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Institution: | (1) Estaci?n Biol?gica de Do?ana, Consejo Superior de Investigaciones Cient?ficas (CSIC), Avda. Am?rico Vespucio s/n, 41092 Sevilla, Spain;(2) Departamento de Biolog?a Experimental, Universidad de Ja?n, Campus Las Lagunillas, s/n, 23071 Ja?n, Spain;(3) Departamento de Biolog?a Animal, Biolog?a Vegetal y Ecolog?a, Universidad de Ja?n, Campus Las Lagunillas, s/n, 23071 Ja?n, Spain |
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Abstract: | A cost-effective, reliable and efficient method of obtaining DNA samples is essential in large-scale genetic analyses. This
study examines the possibility of using a threatened vole species, Microtus cabrerae, as a model for the collection and preservation of faecal samples for subsequent DNA extraction with a protocol based on
the HotSHOT technique. Through the examination of the probability of multi-copies (mitochondrial) and single copy (microsatellite)
loci amplification (including the genotype error) and of the DNA yield (estimated by real-time qPCR), the new protocol was
compared with both the frequently employed methods that successfully use ethanol to preserve faecal samples and with commercial
kit-based DNA extraction. The single-tube HotSHOT-based protocol is a user-friendly, non-polluting, time-saving and inexpensive
method of faeces sample collection, preservation and PCR-quality gDNA preparation. This technique therefore provides researchers
with a new approach that can be employed in high-throughput, noninvasive genetic analyses of wild animal populations. |
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