Comparison of mtDNA extracting methods for common sarcosaphagous insects.
- Author:
Yao-Qing CHEN
1
;
Ya-Dong GUO
;
Mao-Zhi LI
;
Feng XIONG
;
Jian-Bo LI
;
Ji-Feng CAI
Author Information
1. Hunan Police College, Changsha 410138, China. chenyaoqing1994@21cn.com
- Publication Type:Research Support, Non-U.S. Gov't
- MeSH:
Animals;
Coleoptera/genetics*;
DNA Primers;
DNA, Mitochondrial/isolation & purification*;
Diptera/genetics*;
Electrophoresis, Agar Gel;
Entomology;
Forensic Medicine/methods*;
Gene Amplification;
Insecta/genetics*;
Polymerase Chain Reaction/methods*;
Quaternary Ammonium Compounds/chemistry*;
Rabbits;
Reproducibility of Results;
Sequence Analysis, DNA;
Sodium Dodecyl Sulfate/chemistry*
- From:
Journal of Forensic Medicine
2011;27(4):265-270
- CountryChina
- Language:Chinese
-
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.
