Comparison of three different methods extraction of DNA from Apis mellifera meda (Hymenoptera: Apidae, Apis)

Many different methods and technologies are available for the isolation of genomic DNA. The purpose of this study was to test three different method for DNA extraction of Apis mellifera meda based on existing procedures for DNA extraction of plants, fungi and blood. the dominant strain of iran honeybees is Apis mellifera meda and subspescies has several sub-communities throughout the country. In general, all methods involve disruption and lysis of the starting material followed by the removal of proteins and other contaminants and finally recovery of the DNA.Methods: Nurse bees of Apis mellifera meda (< 10 days) from Three colonies (Farm of Khorasgan University,Isfahan) were collected and stored at −80oC until needed. Three different methods were tested: A) Salting-out methods by Miller.S.A. et al.(1988), B)Phenol-chloroform by John,S.W.M. et al. (1991) and C) CTAB (cetyl trimethylammonium bromide) by Murray HG, Thompson WF(1980). Basically, the three methods tested differ in the concentrations of specific components of the extraction buffer. typically, Removal of proteins in three method achieved by digestion with proteinase K. The choice of a method for DNA extraction depends on many factors. Eventually, DNA is separated from cellular components.Results: The results showed that using CTAB and Phenol-chloroform both resulted the best qualitatively and quantitatively in DNA extraction. But after PCR-reaction, The results were different. This may be the remaining phenol extraction DNA. In Salting-out methods, Contamination was high and had to be washed with chloroform. Nevertheless, extraction by the CTAB method was more reliable without contamination or less contamination.

کلیدواژه ها:

DNA extraction ، CTAB ، Phenol-chloroform ، Apis mellifera meda