Comparison of different DNA extraction procedures for Eimeria oocysts

Abstract

A study was carried out to compare the efficacy of different DNA extraction procedures for Eimerian oocysts. A total of 753 litter and fecal samples and intestines were collected from desi poultry farms and slaughter houses in Bengaluru, Karnataka. Out of five different methods of DNA extraction procedures the glass bead method was found to be the best method. Since the above method involves breaking of the oocysts and sporocysts walls and dissolving the sporozoites membrane. This greatly simplifies the currently used DNA extraction procedures for Eimeria species. The liquid nitrogen method was also found to be good but was more critical and time consuming. Other methods such as sodium hypochlorite, direct and sonication has failed to yield DNA from Eimeria oocysts. The present study indicates that glass beads method yielded better quality of DNA for molecular analysis and it is suitable for both large and small number (˂1x103) of oocysts in a sample. Hence, can be used as a routine procedure for DNA extraction for Eimerian oocysts.