DNA extraction and barcode identification of development stages of forensically important flies in the Czech Republic
Language English Country England, Great Britain Media print-electronic
Document type Comparative Study, Journal Article, Research Support, Non-U.S. Gov't
- Keywords
- Forensic science, cytochrome oxidase subunit I, forensic entomology, molecular markers, species identification,
- MeSH
- Diptera classification genetics growth & development MeSH
- DNA isolation & purification MeSH
- Insect Proteins genetics MeSH
- Reagent Kits, Diagnostic MeSH
- Electron Transport Complex IV genetics MeSH
- Sequence Analysis, DNA MeSH
- Forensic Genetics methods MeSH
- DNA Barcoding, Taxonomic methods MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Comparative Study MeSH
- Geographicals
- Czech Republic MeSH
- Names of Substances
- DNA MeSH
- Insect Proteins MeSH
- Reagent Kits, Diagnostic MeSH
- Electron Transport Complex IV 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.
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