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DNA extraction and barcode identification of development stages of forensically important flies in the Czech Republic
T. Olekšáková, M. Žurovcová, V. Klimešová, M. Barták, H. Šuláková,
Jazyk angličtina Země Anglie, Velká Británie
Typ dokumentu srovnávací studie, časopisecké články, práce podpořená grantem
- MeSH
- Diptera klasifikace genetika růst a vývoj MeSH
- DNA izolace a purifikace MeSH
- hmyzí proteiny genetika MeSH
- reagenční diagnostické soupravy MeSH
- respirační komplex IV genetika MeSH
- sekvenční analýza DNA MeSH
- soudní genetika metody MeSH
- taxonomické DNA čárové kódování metody MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
- Geografické názvy
- Česká republika MeSH
Several methods of DNA extraction, coupled with 'DNA barcoding' species identification, were compared using specimens from early developmental stages of forensically important flies from the Calliphoridae and Sarcophagidae families. DNA was extracted at three immature stages - eggs, the first instar larvae, and empty pupal cases (puparia) - using four different extraction methods, namely, one simple 'homemade' extraction buffer protocol and three commercial kits. The extraction conditions, including the amount of proteinase K and incubation times, were optimized. The simple extraction buffer method was successful for half of the eggs and for the first instar larval samples. The DNA Lego Kit and DEP-25 DNA Extraction Kit were useful for DNA extractions from the first instar larvae samples, and the DNA Lego Kit was also successful regarding the extraction from eggs. The QIAamp DNA mini kit was the most effective; the extraction was successful with regard to all sample types - eggs, larvae, and pupari.
Citace poskytuje Crossref.org
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