Comparison of mtDNA extracting methods for common sarcosaphagous insects / 法医学杂志
Journal of Forensic Medicine
; (6): 265-270, 2011.
Article
em Zh
| WPRIM
| ID: wpr-983662
Biblioteca responsável:
WPRO
ABSTRACT
OBJECTIVE@#To compare effects of three different methods for mtDNA extraction from common sarcosaphagous insects including cetyl trimethyl ammonium bromide (CTAB) method, sodium dodecyl sulfate-potassium acetate (SDS-KAc) method and sodium dodecyl sulfate-proteinase K (SDS-PK) method.@*METHODS@#Seventy-two insects from four species [Chrysomya megacephala (Fabricius, 1784), Eusilpha bicolor (Fairmaire, 1896), Paraeutrichopus pecoudi (Mateu, 1954), Vespa velutina (Lepeletier, 1836)] were collected from the corpses of the rabbits in Changsha district. The total DNA of above samples was extracted by CTAB, SDS-Kac and SDS-PK methods. The purity and concentration of DNA were examined by protein-nucleic acid spectrophotometry, and mtDNA were amplified by specific primers and PCR products were detected by agarose gel electrophoresis. Then PCR products were sequenced and subsequently up-loaded to GenBank.@*RESULTS@#mtDNA was successfully extracted with three methods from most of the samples. The SDS-PK method was better in DNA purity compared to other methods and the CTAB method was superior in extracting DNA from old samples, while SDS-KAc method showed no significant difference for extraction effects of different samples.@*CONCLUSION@#The most appropriate method should be chosen depending on different situations. SDS-PK method is expected to obtain high-quality DNA, while CTAB method is preferred in extracting obsolete samples. SDS-KAc method is low cost and can be used in various kinds of preliminary experiments.
Texto completo:
1
Índice:
WPRIM
Assunto principal:
Dodecilsulfato de Sódio
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Besouros
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DNA Mitocondrial
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Amplificação de Genes
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Reação em Cadeia da Polimerase
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Reprodutibilidade dos Testes
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Análise de Sequência de DNA
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Primers do DNA
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Dípteros
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Eletroforese em Gel de Ágar
Limite:
Animals
Idioma:
Zh
Revista:
Journal of Forensic Medicine
Ano de publicação:
2011
Tipo de documento:
Article