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

Alasaad, Samer; Sánchez, Antonio; García-Mudarra, Juan L.; Jowers, Michael J.; Pérez, Jesús M.; Marchal, Juan A.; Romero, Ismael; Garrido-García, José A.; Soriguer, Ramón C.

doi:10.1007/s10344-011-0526-x

Cited by 4 publications

(1 citation statement)

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“…DNA extraction methods that use brief boiling of samples at high temperatures reduce time, labor and cost and have been demonstrated as an efficient DNA extraction technique in many studies (Kieleczawa, 2006;Alasaad et al, 2012;Kitpipit et al, 2014b). DNA extraction by boiling samples in alkaline lysis (AL) buffers has been successfully applied to amplify DNA samples from blood, feathers and many other tissues (Truett et al, 2000;Haunshi et al, 2008;Girish et al, 2013), but the different modifications/compositions of AL buffers have never been compared for their DNA extraction potential to identify vertebrate species in meat and meat products.…”

Section: Introductionmentioning

confidence: 99%

Comparison of different DNA preparatory methods for development of a universal direct PCR-RFLP workflow for identification of meat origin in food products

et al. 2023

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The quality and quantity of the extracted DNA are two key aspects for a successful PCR (Polymerase Chain Reaction) amplification. Moreover, reduction in time and cost required for DNA extraction are also two considerable factors, in cases when large number of samples are to be analyzed within a limited time-frame and budget. Accordingly, the aim of this study was to compare and optimize performance of five different DNA extraction methods by boiling meat tissues from cattle, buffalo, sheep, goat, chicken, camel, horse and dog in PBS (Phosphate Buffer Saline), distilled water, alkaline lysis buffers 1, 2 or 3. The results indicated that the boiling of meat and its products in alkaline lysis buffers was a good method to extract crude DNA. The optimized crude DNA extraction protocol was coupled with PCR-RFLP (Restriction Fragment Length Polymorphism) analysis for meat species differentiation. This developed workflow was tested on fifty-three commercial beef and mutton samples, out of which three samples were found to be adulterated. In conclusion, the rapid crude DNA extraction protocol has led to the development of a direct PCR-RFLP workflow that is simple, time-saving and cost-effective for PCR-based identification of different meat species.

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Section: Introductionmentioning

confidence: 99%

Comparison of different DNA preparatory methods for development of a universal direct PCR-RFLP workflow for identification of meat origin in food products

et al. 2023

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Development and evaluation of hotshot protocols for cost- and time-effective extraction of PCR-ready DNA from single freshwater mussel larvae (Bivalvia: Unionida)

Zieritz

Yasaeng

Razak

et al. 2018

Journal of Molluscan Studies

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The use of radio-collars for monitoring wildlife diseases: a case study from Iberian ibex affected by Sarcoptes scabiei in Sierra Nevada, Spain

et al. 2013

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BackgroundWildlife radio tracking has gained popularity during the recent past. Ecologists and conservationists use radio-collars for different purposes: animal movement monitoring, home range, productivity, population estimation, behaviour, habitat use, survival, and predator-prey interaction, among others. The aim of our present study is to highlight the application of radio-collars for wildlife diseases monitoring. The spread of wildlife diseases and the efficacy of management actions for controlling them propose serious challenges for ecologists and conservationists, since it is difficult to re-capture (or simply observe) the same animal in pre-determined temporal interval, but such difficulty is overcome by the use of gps-gsm radio collars.MethodsIn the present study we report, for the first time to our knowledge, the use of radio-collars in the monitoring of Iberian ibex affected by Sarcoptes scabiei in Sierra Nevada mountain range, Spain. Twenty-five moderate or slightly mangy animals were radio-collared between 2006 and 2013.ResultsThe radio-collars allowed us to confirm the presence of resistance to S. scabiei within Iberian ibex population. Twenty (80%) of the collared animals recovered totally from mange, while the disease progressed in the other five Iberian ibex (20% of the collared animals) and the animals died. The average estimated recovery time of the resistant animals was 245 ± 277 days, and the estimated average survival time of the non-resistant Iberian ibex was 121 ± 71 days. Non-resistant animals survived at least 100 days, while all of them died with less than 200 days. Sixty per cent of the resistant animals were recovered with less than 200 days.ConclusionsWe report, for the first time, the successful use of radio collars for wildlife diseases monitoring using Iberian ibex/S. scabiei as a model. By using radio collars we documented that most of the Sarcoptes-infected Iberian ibex are resistant to this disease, and we estimated the average time for Iberian ibex recovering from mange infection and the average survival time of the non-resistant ones. We expect wider use of radio-collars for wild animals diseases monitoring, affected/not-affected animals interaction, and treatment efficacy, among others.

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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

Cited by 4 publications

References 24 publications

Comparison of different DNA preparatory methods for development of a universal direct PCR-RFLP workflow for identification of meat origin in food products

Comparison of different DNA preparatory methods for development of a universal direct PCR-RFLP workflow for identification of meat origin in food products

Development and evaluation of hotshot protocols for cost- and time-effective extraction of PCR-ready DNA from single freshwater mussel larvae (Bivalvia: Unionida)

The use of radio-collars for monitoring wildlife diseases: a case study from Iberian ibex affected by Sarcoptes scabiei in Sierra Nevada, Spain

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