Modification of a CTAB DNA extraction protocol for plants containing high polysaccharide and polyphenol components
Modification of a CTAB DNA extraction protocol for plants containing high polysaccharide and polyphenol components
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DOI：
10.1007/bf02772108
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发表时间：
1997-03-01
影响因子：
2.1
通讯作者：
Baum, BR
中科院分区：
文献类型：
--
作者：
Porebski, S;Bailey, LG;Baum, BR
描述了一种相对快速、廉价且一致的从含有大量多酚、单宁和多糖的展开叶材料中提取 DNA 的方案。成熟的草莓叶含有高含量的这些次要成分，被用作研究组。该方法包括改进的 CTAB 提取、采用高盐浓度去除多糖、使用聚乙烯吡咯烷酮 (PVP) 去除多酚、延长 RNase 处理以及苯酚-氯仿提取。野生和栽培的八倍体和二倍体草莓物种的平均产量范围为 20 至 84 微克/克成熟叶组织。对 60 株植物的结果进行了检查，结果在 RAPD 反应中始终可以扩增，每 25 微升反应只需 0.5 ng DNA。目前，这是从成熟草莓叶组织中分离 DNA 的第一个程序，该程序对各种不同物种（八倍体和二倍体）产生一致的结果，并且在长期储存之前和之后都是稳定的且可进行 PCR 扩增。该程序可能对蔷薇科中的其他困难物种有用。
A relatively quick, inexpensive and consistent protocol for extraction of DNA from expanded leaf material containing large quantities of polyphenols, tannins and polysaccharides is described. Mature strawberry leaves, which contain high levels of these secondary components, were used as a study group. The method involves a modified CTAB extraction, employing high salt concentrations to remove polysaccharides, the use of polyvinyl pyrrolidone (PVP) to remove polyphenols, an extended RNase treatment and a phenol-chloroform extraction. Average yields range from 20 to 84 mu g/g mature leaf tissue for both wild and cultivated octoploid and diploid Fragaria species. Results from 60 plants were examined, and were consistently amplifiable in the RAPD reaction with as little as 0.5 ng DNA per 25-mu L reaction. Presently, this is the first procedure for the isolation of DNA from mature strawberry leaf tissue that produces consistent results for a variety of different species, both octoploid and diploid, and is both stable and PCR amplifiable before and after extended storage. This procedure may prove useful for other difficult species in the family Rosaceae.