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Single-tube HotSHOT technique for the collection, preservation and PCR-ready DNA preparation of faecal samples: the threatened Cabrera's vole as a model

AuthorsAlasaad, Samer ; Sánchez, Antonio; García-Mudarra, Juan L. ; Jowers, Michael J.; Pérez, Jesús M.; Marchal, Juan Alberto; Romero, Ismael; Garrido-García, José A.; Soriguer, Ramón C.
KeywordsNoninvasive method
Sample collection
Sample preservation
DNA extraction
HotSHOT technique
Kit-based protocol
Real-time qPCR
Issue Date2012
CitationEur J Wildl Res (2012) 58:345–350
AbstractA 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
Publisher version (URL)http://dx.doi.org/10.1007/s10344-011-0526-x
Appears in Collections:(EBD) Artículos
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