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梨不同DNA提取方法的效果研究
引用本文:乌云塔娜,张党权,谭晓风.梨不同DNA提取方法的效果研究[J].中国生物工程杂志,2003,23(7):98-101.
作者姓名:乌云塔娜  张党权  谭晓风
作者单位:1. 中南林学院经济林育种与栽培国家林业局重点实验室, 株洲412006; 2. 中山大学教育部基因工程重点实验室, 广州510275
基金项目:湖南省科技三项基金资助项目 ( 0 0JZY2 15 5 )
摘    要:以7个梨品种为实验材料,比较分析了SDS法、CTAB法、SDSCTAB法、改良的CTAB法、高盐低pH值法、分步离心法对梨总DNA提取的效果。结果表明:利用以上6种方法提取的梨总DNA在纯度和量上有很大的差别。所得到的平均DNA量从大到小依次为:分步离心法、SDS法、SDSCTAB法、改良的CTAB法、CTAB法、高盐低pH值法。DNA提取纯度依次为分步离心法、SDSCTAB法、改良的CTAB法、高盐低pH值法、CTAB法、SDS法。RAPD和自交不亲和基因(S基因)特异性引物扩增实验结果都比较理想,但分步离心法和SDSCTAB法提取的DNA双酶切效果较好。分步离心法提取的梨总DNA更适用于后续的分子生物学实验操作。

关 键 词:  DNA提取  RAPD  自交不亲和  

Studies on Methods of Isolating Pear DNA
Wuyun Tana,Zhang Dangquan,Tan Xiaofeng.Studies on Methods of Isolating Pear DNA[J].China Biotechnology,2003,23(7):98-101.
Authors:Wuyun Tana  Zhang Dangquan  Tan Xiaofeng
Institution:Wuyun Tana1 Zhang Dangquan2 Tan Xiaofeng1
Abstract:Effects on isolating pear genomic DNA by using SDS,CTAB,SDS-CTAB,improved CTAB,high salt-low pH and fractional centrifugation are analyzed and compared.Seven varieties of pear are used.The result shows that the purity and yield of pear DNA obtained by above methods is distinctly different.These six methods are ranked as:fractional centrifugation,SDS,SDS-CTAB,improved CTAB,CTAB and high salt-low pH according to the average yields of pear DNA in mass from high to low,and ranked as:fractional centrifugation,SDS-CTAB,improved CTAB,high salt-low pH,CTAB and SDS according to the pear DNA purities from high to low.Above pear DNA obtained by these six methods was subjected to RAPD analyses and works well,and DNA obtained by methods of fractional centrifugation and SDS-CTAB works better than others for double enzymatic digestion.These results suggest that pear DNA obtained by fractional centrifugation display superiority than others for molecular biological operations.
Keywords:Pear  DNA isolation  RAPD  Self-incompatibility
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