高质量丝栗栲基因组DNA的提取方法

为了促进丝栗栲分子生物学的研究,采用尿素法、柠檬酸钠法、改良CTAB法和SDS法4种方法提取丝栗栲叶片基因组DNA,并对提取效果进行了比较和分析。结果表明,改良CTAB法较适合丝栗栲基因组DNA提取,其提取的DNA产率高,凝胶检测条带整齐清晰,杂质少、无明显降解,OD260/OD280比值在1.8左右;而尿素法难以提取基因组DNA,SDS法提取的DNA得率较低,柠檬酸钠法有蛋白质、多糖等杂质污染均不太适用丝栗栲基因组DNA提取。采用改良CTAB法提取得到丝栗栲幼叶、老叶、嫩芽和幼茎基因组DNA,产率分别为177.3、167.1、242.0和156.7μg/g。酶切分析也证明,改良CTAB法提取的不同组织DNA适用于进行后续分子生物学研究。

Extraction method of high quality DNA for Castanopsis fargesii Franch

In order to accelerate the molecular biology research of Castanopsis fargesii,DNA of C.fargesii was extracted from the fresh leaves by carbamide method,sodium citrate method,improved CTAB and SDS method separately,and the extraction effects were compared in this study.The results showed that the improved CTAB method was suitable for extracting the genomic DNA from C.fargesii.DNA quality was high,degradation was light and impurity was little,and OD260/OD280 was 1.80 or so.However,the genomic DNA could not be extracted by carbamide method.The yield of DNA sample extracted by SDS method was very low.Sodium citrate method was difficult to get rid of polyhexose completely,DNA browned and degraded seriously.Therefore,Carbamide method,SDS method and Sodium citrate method were not suitable for extracting the genomic DNA from C.fargesii.The yield of DNA of young leaves,old leaves,young shoots and young stems obtained by the improved CTAB method was 177.3,167.1,242.0 and 156.7 μg/g separately.When tested through the restriction enzyme analysis,the DNA samples extracted from different tissues using improved CTAB method could meet the next experiment requirement perfectly.