【作者】 农向群；

【Author】 NONG Xiangqun (Institute of Biological Control,CAAS,Beijing 100081,China)

【机构】 中国农科院生物防治研究所；

【摘要】 <正>研究比较3种方法提取白僵菌DNA的效果并给予改进。结果表明,在无液氮条件下,可用氯化苄促使细胞破裂,释放DNA,但此法效果不很稳定,容易造成DNA降解断裂或收率很低。SDS和CTAB都能很好地分离析出DNA,后者的收率较高,可达>50μg/100mg菌丝。CsCl对清除多糖更多还原

【Abstract】 The effect of 3 methods of extracting DNA from Beauveria bassiana were compared and improved.The results of the experiments showed that SDS(sodium dodecyl sulfate) and CTAB(cetyl-triethyl-ammonium bromidewere) both are effective for isolate DNA from the fungus.A higher yield of DNA could be obtained by CTAB,which get to 50μg/100mg fresh mycelia.Benzyl chloride could break cells and made them release DNA without being ground in liquid nitrogen.But the protocol tended to cause lower yield and more DNA digestion.Polysacchrides from fungal cell was removed with CsCl.High quality and quantity of DNA from the fungus could be provided by the modified procedure with CTAB,which was practical for isolating DAN from a large number of fungal samples. The extracted DNA was suitable for RAPD-PCR and RFLP-PCR analysis.更多还原