Abstract
The aim of the present study was to assess whether individual Sarcoptes mites collected from frozen skin (‘postponed isolation’ method) are suitable sources of PCR-quality genomic DNA, and to test the effectiveness of this method in comparison with the ‘direct isolation’ method, often used through force of habit. Hundreds of single Sarcoptes scabiei samples, resulting from direct (live) or postponed (post-frozen) isolation, were tested using a ~450 bp product (ITS-2) and multi-locus 10× genotyping with microsatellite markers. No statistical difference in yield of soluble DNA was found between the two isolation methods. Nevertheless, 19% of the reactions were classified as failed preparations in the direct isolation method, whereas the rate of unsuccessful reactions was 34% in the postponed isolation method. Consequently, post-frozen isolation is suitable and recommendable for Sarcoptes mite gDNA preparation, particularly when performing a balancing act among safety, practicability and profitability. These results have implications for mite collection for DNA extraction, and hence the needed wider leap of Sarcoptes into the genetic era.
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Acknowledgements
We would like to thank all the people who supported the Department of Animal Production, Epidemiology and Ecology (University of Turin-Italy) with Sarcoptes samples, and RNM 118 research group (Junta de Andalucía-Spain) for supporting SA’s investigation stay in Italy. The research was supported by MURST contract year 2004, Prot. 2004078701_001 (LR).
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Alasaad, S., Soglia, D., Maione, S. et al. Effectiveness of the postponed isolation (post-frozen isolation) method for PCR-quality Sarcoptes mite gDNA. Exp Appl Acarol 47, 173–178 (2009). https://doi.org/10.1007/s10493-008-9196-0
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DOI: https://doi.org/10.1007/s10493-008-9196-0