DNA extraction and barcode identification of development stages of forensically important flies in the Czech Republic

0473646 - BC 2019 RIV GB eng J - Článek v odborném periodiku
Olekšáková, T. - Žurovcová, Martina - Klimešová, V. - Barták, M. - Šuláková, H.
DNA extraction and barcode identification of development stages of forensically important flies in the Czech Republic.
Mitochondrial DNA Part A. Roč. 29, č. 3 (2018), s. 427-430. ISSN 2470-1394. E-ISSN 2470-1408
Institucionální podpora: RVO:60077344
Klíčová slova: forensic science * forensic entomology * species identification
Obor OECD: Genetics and heredity (medical genetics to be 3)
Impakt faktor: 0.566, rok: 2018

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.
Trvalý link: http://hdl.handle.net/11104/0285084