一种果梅休眠枝韧皮部DNA提取方法  

A method of extracting DNA from the phloem of dormant wood of Prunus mume

作　　者：王飞[1] 沈玉英[1,2] 佟兆国[1] 高志红[1] 章镇[1] 

机构地区：[1]南京农业大学园艺学院,南京210095 [2]浙江建设职业技术学院,杭州311231

出　　处：《上海农业学报》2011年第1期55-59,共5页Acta Agriculturae Shanghai

基　　金：国家自然科学基金(30671438;30871681)资助

摘　　要：以5个果梅品种的休眠枝韧皮部为材料,采用改良CTAB法提取其基因组总DNA,并与其相应幼叶的DNA提取效果进行比较。结果表明:该方法从5个果梅品种的休眠枝韧皮部中均能提取出基因组总DNA,且DNA具有较高的纯度和完整性,5个品种的D_(260nm)/D_(280nm)比值为1.8～2.0,无降解现象,RNA去除干净,能被限制性内切酶完全消化;经SSAP分析验证,韧皮部DNA的电泳表型与其相应幼叶完全一致(引物组合为LTR-1/EcoR-ACC,LTR-2/EcoR-ACC),且均表现为条带清晰、多态性好,表明此方法提取的果梅休眠枝韧皮部总DNA完全适于SSAP分析。The phloems of 5 Prunus mume cultivars＇ dormant woods were used as test materials, their genomic DNAs were extracted by the improved CTAB method,and the effect of DNA extraction from the phloems was contrasted with that from their young leaves.The results showed that all the genomic DNAs extracted from the phloems by this method were rather good in both purity and integrality, the D_（260nm）/D_（280nm） ratio was 1.8～2.0,no degradation occurred,the RNA was eliminated completely, and the DNAs could be completely digested by restriction endonucleases.The SSAP analysis indicated that the electrophoretic phenotype of the phloem DNA was just the same as that of the leaf DNA（the primer combination：LTR-1/EcoR-ACC,LTR-2/Eco R-ACC）,being clear in band and good in polymorphism. It was concluded that the total DNA extracted by this method from the phloem of dormant wood of P.mume was completely suitable for the SSAP analysis.

关 键 词：果梅 DNA提取 韧皮部 SSAP 

分 类 号：Q78[生物学—分子生物学]