摘要
经反复实验,采用在抽提前加入一定量的抗氧化剂β 巯基乙醇,用冰冻的异丙醇来沉淀DNA,并增加氯仿/异戊醇抽提次数的所谓改良CTAB法,对常山胡柚叶片DNA抽提与分析,不仅成功提取到常山胡柚叶片DNA,而且所得的DNA浓度高、纯度好,20个试样的DNA平均质量浓度为272 86μg/mL,平均每克鲜叶片可获DNA约82μg,纯度基本上都在1 4～1 8范围内,并经RAPD反应验证,完全能满足常规分子生物学分析的要求。
Changshan huyou is a new species of citrus native to Changshan County,Zhejiang Province.So far,there is no report at home and abro ad on the extraction and analysis of DNA in its leaf.After repeated experiments, through the improved CTAB method of addi ng a certain amount of anti oxidant β me rcaptoethanol,precipitating DNA by using frozen isopentenol and increasing the t imes of chloroform/isopentyl alcohol ext racting,we succeed in extracting DNA of high density and purity of in the leaf o f changshan huyou.The average density of 20 samples is 272 86 μg/mL.And about 82 μg DNA can be obtained from per grams fresh leaf.The purity ranges from 1 4 to 1 8,which fully meets the demand of norm al molecular biological analysis through RAPD response test.
出处
《江苏林业科技》
2003年第1期5-8,共4页
Journal of Jiangsu Forestry Science & Technology
基金
浙江省教育厅基金资助项目(97128)
关键词
常山胡柚
叶片
DNA抽提
改良CTAB法
Changshan huyou(Hush addock)
Leaf
DNA extraction
Improved CTA B method
