摘要
用氯化锂法从眼点拟微球藻 (Nannochloropisisoculata)中提取DNA并对其提取条件进行了优化。新鲜藻细胞在提取液 (0.6mol/LLiCl,0.8%Sakosyl,20mmol/LEDTA(pH8.0) ,0.4%PVP,10mmol/LTris HCl(pH8.0) ,5% β 巯基乙醇 )中经过55℃温育30min,每mg鲜藻可提取20～40ngDNA ,提取的DNA分子量在23kb左右 ,可用于PCR、限制性酶切等。
A rapid and simple method for DNA extraction frommicroalgae Nannochloropisis oculata using LiCl has been developed and optimized.Treating fresh algae in a solution containing0.6mol/L LiCl,0.8%Sakosyl,20mmol/L EDTA(pH8.0),0.4%PVP,10mmol/LTrisˉHCl(pH8.0),5%β-mercaptoethanol at55℃for30minutes can extract20～40μg DNA from one gram fresh algae.The DNA is qualitified as the PCR template amplification and the substrate of Restriction Endonu-clease.The DNA is about23kb in length deter mined by agarose gel assays.
出处
《海洋科学》
CAS
CSCD
北大核心
2003年第9期58-60,77,共4页
Marine Sciences
基金
国家"863"计划项目819 02 01号
