摘要
苎麻组织中含有较多的酚类、粘状物及黄酮类等次生代谢物质,其mtDNA的提取比较困难。本实验中采用蔗糖衬垫法提取苎麻mtDNA。提取过程中往缓冲液中加入抗氧化性物质来减轻DNA的褐变;同时在酚-氯仿/异戊醇抽提过程中加入高盐溶液,用以去除多糖;且采用擦镜纸而不是纱布过滤可提高mtDNA的得率。mtDNA的ISSR检测结果表明,蔗糖衬垫法提取mtDNA时,辅助加入抗氧化剂和高盐溶液效果较好。
The extraction of mtDNA was difficult than that of gDNA,due to a high content of secondary metabolic products such as polysaccharides,hydroxybenzene and flavone in ramie [Boehmeria nivea（L.） Gaud],an optimized protocol for ramie mtDNA extraction based on sucrose-mediated sedimentation was proposed.Antioxidant was added into the extraction buffer,which could prevent the solution from browning.High concentration salt solution of 4 % NaCl was used to remove polysaccharides in the extraction process with chloroform-isopentanol.The mtDNA templates extracted with the above modified method presented better band patterns in ISSR-PCR amplification,indicating that the combination of sucrose-mediated sedimentation method with antioxidant and high salt solution was suitable for the extraction of ramie mtDNA.
出处
《西南农业学报》
CSCD
北大核心
2010年第4期1377-1379,共3页
Southwest China Journal of Agricultural Sciences
基金
国家自然科学基金项目(30971825)
关键词
苎麻
mtDNA提取
蔗糖衬垫法
改良
Ramie
mtDNA extraction
Sucrose-mediated sedimentation
Modified protocol
