Comparative study of extraction methods for genomic DNA of Cryptococcus neoformans

Objective To establish simple and rapid methods for extracting genomic DNA from C.neoformans.To compare and search the better one for gene analysis.Methods Glass beads method,classical lytic enzyme treatment,3%SDS lysis and the benzyl chloride method were set up and compared.In glass beads method,cells were broken by vigorously mixing with glass beads on a votorex;In 3%SDS method,cells were heated in 3%SDS extraction buffer containing 10 mmol/L DTT for 20 minutes.The lysate was extracted with 5 mmol/L KAc and isopropanol.Yield of DNA was calculated from the A260 for clean DNA.Results DNA production by 3%SDS lysis,classical lytic enzyme treatment,glass beads method and the benzyl chloride method was 0.415 4±0.036 7,0.848 4±0.075 6,1.263 6±0.204 0,0.407 0±0.033 9 (g/L×108 CFU/mL) respectively.Conclusions Glass beads method was more sensitve,reproducible,simple and cost-effective for DNA extraction.