摘要
目的 探讨几种快速提取新生隐球菌DNA方法的效果。方法 分别用玻璃珠机械破裂法、酶消化法、高渗剂破裂法及三法联合法提取有荚膜的新生隐球菌B 3 50 1野生株DNA ,用联合法提取 5种血清型新生隐球菌标准株的DNA ,用紫外分光光度计检测提取DNA的浓度和纯度。结果 4种方法抽提 2ml新生隐球菌培养液 (菌体数约为 1×10 8个 /ml) ,可获得DNA的量约是 64~ 2 0 3 μg。酶消化法、高渗剂破裂法及三法联合法抽提的DNA ,其OD2 60 /OD2 80比值均小于 1.8。结论 除玻璃珠机械破裂法外 ,酶消化法、高渗剂破裂法 ,以及三法联合法均可用于快速抽提新生隐球菌的DNA。
Objective To evaluate some rapid methods of e xtracting DNA from Cryptococcus neoformans.Methods DNA from the encapsulated wilde-typed isolates B3501 of Cryptococcus neoformans was extracted by glass beads mechanical disru ption method,enzyme digestion method,chaotropic agents disruption method and com bined method.DNA from five serotype standard strains of Cryptococcus neoformans were extracted only by combined method.The concentration and purity of DNA ext raceted was detected with UV spectrophotometer.Results Yields of 64-203 μg of total DNA from 2 ml cultu re(1×10 8 cells/ml) were obtained.The ratios OD 260/OD 280 of DNA extracted wi th each of the later three methods was less than 1.8.Conclusion Except glass beads mechanical disruption metho d,the other methods are suitable for rapid extracting DNA from Cryptococcus neof ormans.The combined method is the most optimal method.
出处
《中国皮肤性病学杂志》
CAS
北大核心
2003年第6期410-411,共2页
The Chinese Journal of Dermatovenereology