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  • [Comparison of mtDNA extracting methods for common sarcosaphagous insects].

[Comparison of mtDNA extracting methods for common sarcosaphagous insects].

Fa yi xue za zhi (2011-09-15)
Yao-Qing Chen, Ya-Dong Guo, Mao-Zhi Li, Feng Xiong, Jian-Bo Li, Ji-Feng Cai
ABSTRACT

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. 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. 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. 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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Ammonium lauryl sulfate solution, ~30% in H2O
Sigma-Aldrich
Ammonium lauryl sulfate solution, ~30% in H2O (T)
Sigma-Aldrich
Lithium dodecyl sulfate, BioXtra, ≥98.5% (GC)
Sigma-Aldrich
Lithium dodecyl sulfate, BioReagent, for molecular biology, suitable for electrophoresis
Sigma-Aldrich
Lithium dodecyl sulfate, ≥98.5% (GC)