文献类型：期刊文献

中文题名：KingFisher Flex核酸纯化系统上树木叶片DNA的提取方法

英文题名：A Method for Using KingFisher Flex Purification System to Isolate DNA from Tree Leaves

作者：杨合宇[1,2] 李发根[2] 翁启杰[2] 李梅[2] 陈升侃[2] 姬红霞[2] 周长品[2] 甘四明[1,2]

第一作者：杨合宇

机构：[1]中国林业科学研究院林木遗传育种国家重点实验室,北京100093;[2]中国林业科学研究院热带林业研究所,热带林业研究国家林业局重点实验室,广州510520

年份：2017

卷号：15

期号：5

起止页码：1854-1861

中文期刊名：分子植物育种

外文期刊名：Molecular Plant Breeding

收录：CSTPCD;;北大核心:【北大核心2014】；CSCD:【CSCD2017_2018】；

基金：广东省自然科学基金(博士启动:2016A030310007);广东省林业科技创新项目(2016KJCX014);中国林科院基本科研业务费专项(CAFYBB2017SY018)共同资助

语种：中文

中文关键词：树木;DNA提取;KingFisher;Flex

外文关键词：Tree, DNA isolation, KingFisher flex

分类号：S

摘要：以桉树4种/杂种的叶片为材料,比较了5种磁珠试剂盒在King Fisher Flex核酸纯化系统上自动提取的DNA得率和纯度,初步选出了2种较好的试剂盒。比较了King Fisher Flex上不同的洗涤速度,最优为Medium。初选的2种试剂盒所提的DNA均可有效用于PCR扩增,表明DNA纯度均较好,确定得率最高的为最优试剂盒。对比手动的改良CTAB法(DNA得率19.0μg/g,OD_(260)/OD_(280)为1.97,OD_(260)/OD_(230)为2.03),本研究优化的方法可显著提高DNA得率(103.6μg/g)且DNA纯度相似(OD_(260)/OD_(280)为1.93,OD_(260)/OD_(230)为1.89)。优化的方法可有效用于马占相思、银中杨、降香黄檀、思茅松、马尾松和短枝木麻黄叶片的DNA提取。这对树木分子生物学研究的高通量DNA提取有重要的应用价值,对非木本植物的DNA提取和其他基于磁珠法的纯化系统亦可提供有益的方法参考。
In this research, five magnetic DNA kits were tested for yield and purity of DNA isolated from leaves of four Eucalyptus species/hybrids by using KingFisher Flex purification system, and two superior kits were preliminarily selected out. The washing speed ＇Medium＇ was determined from the comparison of four settings on KingFisher Flex. DNA samples isolated from the two superior kits could result in effective PCR amplification, which indicated good DNA purity, and the highest yielding kit was the choice of our method. Compared with manually performing CTAB method （DNA yield 19.0 ixg/g, OD26o/OD28o 1.97 and ODz6o/OD23o 2.03）, our method could increase considerably the DNA yield （103.6 μg/g） while similar DNA purity （OD260/OD280 1.93 and 0D260/ OD230 1.89） was retained. Our method was effectively applied to isolate DNA from leaves of Acacia mangium, Populus albaxPo, berolinensis, Dalbergia odorifera, Pinus kesiya var. langbianensis, Pi. massoniana and Casuarina eqtdsetifolia. This could be valuable for high throughput DNA isolation in tree molecular biology studies and would provide potential method for isolating DNA from non-woody plants and/or with other mangetic particle based purification systems.