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A Noninvasive Hair Sampling Technique to Obtain High Quality DNA from Elusive Small Mammals
Published on: March 13, 2011
Two simple techniques for the safe Sarcoptes collection and individual mite DNA extraction
Dominga Soglia1, Luisa Rambozzi, Sandra Maione
1Dipartimento di Produzioni Animali, Epidemiologia ed Ecologia, Università degli Studi di Torino, 10095 Grugliasco, Italy. dominga.soglia@unito.it
Abstract:
Availability of mites is a recognized limiting factor of biological and genetic investigations of the genus Sarcoptes. Current methods of deoxyribonucleic acid (DNA) extraction from individual mites also need substantial improvement in efficiency and operator friendliness. We have first developed a technique for efficient and safe extraction of living mites from scabietic skin samples (crusts or deep skin scrapings). Its core device is a large plastic syringe connected with a 1.5-ml Eppendorf tube. The source material is introduced in the syringe and the device in a shoe box with the tip half of the tube emerging. Mites migrate towards a heat source during a minimum of 36 h. Then, the tube is detached and the mites utilized without risks for the operators. A second technique allows operator-friendly manipulation of individual mites for DNA extraction. Fixed mites are isolated by adhesion to a small strip of polyvinyl chloride (PVC) adhesive tape operated with tweezers. Then, mite and strip are plunged in the lyses buffer and the sample twice submitted to thermal shock for disruption of the chitinous exoskeleton. Data show that the tape does not interfere with successive DNA extraction with a commercial kit. The corresponding protocol, that we briefly name "PVC adhesive tape + thermal shock + kit DNA extraction," compares favorably with the available ones.
Insights
New methods improve the availability of Sarcoptes scabiei mites for research. These techniques enhance the efficiency and safety of extracting mites and their deoxyribonucleic acid (DNA) from skin samples.
Area of Science:
- Parasitology
- Molecular Biology
- Dermatology
Background:
- Availability of Sarcoptes mites is a major limitation in biological and genetic research.
- Current deoxyribonucleic acid (DNA) extraction methods for individual mites are inefficient and difficult to handle.
Purpose of the Study:
- To develop improved techniques for extracting living Sarcoptes mites from skin samples.
- To create a more user-friendly method for DNA extraction from individual mites.
Main Methods:
- A novel extraction device using a plastic syringe and Eppendorf tube to isolate mites via thermotaxis.
- A polyvinyl chloride (PVC) adhesive tape method for manipulating individual mites for DNA extraction, followed by thermal shock and commercial kit processing.
Main Results:
- The thermotaxis method efficiently and safely isolates living mites from scabietic skin samples.
- The PVC tape method allows for operator-friendly manipulation and does not interfere with subsequent DNA extraction.
- The combined "PVC adhesive tape + thermal shock + kit DNA extraction" protocol is effective and superior to existing methods.
Conclusions:
- The developed methods significantly enhance the accessibility of Sarcoptes mites and their DNA for research.
- These techniques address key limitations in mite availability and DNA extraction, facilitating future biological and genetic investigations.
