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Mol Biol Rep ; 46(6): 6625-6628, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31559510

RESUMO

The novel dipstick DNA extraction method was tested for its reliability and usability for biological barcoding in comparison to a commercial kit and to a simplified isopropanol precipitation method using crayfish gill tissue. Following DNA extraction, the mitochondrial COI-gene was amplified in a PCR-reaction using a standard set of universal invertebrate primers. All three extraction techniques resulted in successful amplifications. With the dipstick method, PCR immediately follows the very brief DNA extraction technique. We suggest that the dipstick method is an affordable, efficient, and reliable DNA extraction method uniquely suited for biological barcoding that results in reliable and reproducible amplification for downstream applications such as sequencing. Additional tests on crayfish with primers for different parts of the mitochondrial genome and on fish using specific fish COI-primers confirmed these findings. Due to the few steps involved in the DNA extraction procedure the dipstick technique is also highly recommended for high school and university biology courses.


Assuntos
Astacoidea/classificação , Código de Barras de DNA Taxonômico/veterinária , DNA Mitocondrial/isolamento & purificação , Animais , Astacoidea/genética , Complexo IV da Cadeia de Transporte de Elétrons/genética , Filogenia , Reação em Cadeia da Polimerase/veterinária , Reprodutibilidade dos Testes
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