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火龙果茎基因组DNA提取方法改良
引用本文:农全东,张明永,张美,简曙光,陆宏芳,夏快飞,文和明.火龙果茎基因组DNA提取方法改良[J].植物学报,2019,54(3):371-377.
作者姓名:农全东  张明永  张美  简曙光  陆宏芳  夏快飞  文和明
作者单位:1. 中国科学院华南植物园, 中国科学院华南农业植物分子分析与遗传改良重点实验室, 广州 5106502. 中国科学院华南植物园, 广东省应用植物学重点实验室, 广州 5106503. 文山州农业科学院, 文山 663099
基金项目:中国科学院A类战略性先导科技专项(XDA13020500和广州市科技计划No.201804010409)
摘    要:火龙果(Hylocereus undulatus)是近年发展起来的一种新兴热带水果, 其茎富含多糖、多酚及其它次生代谢物, 黏性极大, 很难从中提取高质量的DNA。特别是一年生以上的老茎, 目前尚未有较好的DNA提取方法。为了解决这一难题, 该研究对CTAB+Tris-HCl洗涤法进行了3种方式的改良。结果表明, “改进三”方法可不受取样时期和取样部位的限制, 从一年生以上火龙果茎中提取的DNA质量最好且不含黏性物质, 可用于酶切与分子标记等生化和分子生物学实验。该研究探索了一条较为理想的火龙果茎DNA提取方法, 值得推广应用。

关 键 词:火龙果    DNA提取  改进CTAB+Tris-HCl洗涤法  
收稿时间:2018-06-11

Improvement in Extraction Methods of Genomic DNA from Pitaya Stems
Quandong Nong,Mingyong Zhang,Mei Zhang,Shuguang Jian,Hongfang Lu,Kuaifei Xia,Heming Wen.Improvement in Extraction Methods of Genomic DNA from Pitaya Stems[J].Bulletin of Botany,2019,54(3):371-377.
Authors:Quandong Nong  Mingyong Zhang  Mei Zhang  Shuguang Jian  Hongfang Lu  Kuaifei Xia  Heming Wen
Institution:1. Key Laboratory of South China Agricultural Plant Molecular Analysis and Genetic Improvement, South China Botanical Garden, CAS, Guangzhou 510650, China2. Guangdong Provincial Key Laboratory of Applied Botany, South China Botanical Garden, CAS, Guangzhou 510650, China3. Wenshan Academy of Agricultural Sciences, Wenshan 663099, China
Abstract:Pitaya (Hylocereus undulatus) is a tropical fruit developed in recent years. Because its stem is full of polysaccharides, polyphenols and other secondary metabolites, extracting high-quality DNA from the stem is difficult, especially from stems more than 1 year old. At present, there is no good method for DNA extraction. We examined three methods to improve DNA extraction based on the CTAB+Tris-HCl washing method for better and easier DNA extraction. The third method obtained the best-quality DNA without sticky substances from pitaya stem more than 1 year old and was not restricted by sampling data and sampling site. The extracted DNA can be used in enzyme digestion and inter-simple sequence repeat amplification experiments. Our study provides an ideal method for extracting DNA from pitaya stem, which is worth popularizing.
Keywords:pitaya  stem  DNA extraction  improved CTAB+Tris-HCl washing method  
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