Extraction of high-quality genomic DNA from latex-containing plants
Extraction of high-quality genomic DNA from latex-containing plants
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DOI：
10.1016/s0003-2697(02)00665-6
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发表时间：
2003-04-01
影响因子：
2.9
通讯作者：
Van Laere, A
中科院分区：
文献类型：
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作者：
Michiels, A;Van den Ende, W;Van Laere, A
完整的高分子量基因组DNA的分离对于许多分子生物学应用是必不可少的，包括长PCR、内切酶限制性消化、Southern印迹分析和基因组文库构建。许多方案可用于从植物材料中提取DNA。然而，对于含胶乳的菊科（菊苣亚科）物种，标准方案和市售试剂盒不能产生高质量可扩增DNA的有效产量。一个十六烷基三甲基溴化铵协议已被优化的基因组DNA从含乳胶植物的分离。改良方案中的关键步骤是使用黄化叶组织进行提取和过夜25 ℃异丙醇沉淀步骤。纯化的DNA具有优异的光谱质量，被限制性内切酶有效地消化，并且适合于长片段PCR扩增。(C)2003 Elsevier Science（美国）。All rights reserved.
The isolation of intact, high-molecular-mass genomic DNA is essential for many molecular biology applications including long PCR, endonuclease restriction digestion, Southern blot analysis, and genomic library construction. Many protocols are available for the extraction of DNA from plant material. However, for latex-containing Asteraceae (Cichorioideae) species, standard protocols and commercially available kits do not produce efficient yields of high-quality amplifiable DNA. A cetyltrimethylammonium bromide protocol has been optimized for isolation of genomic DNA from latex-containing plants. Key steps in the modified protocol are the use of etiolated leaf tissue for extraction and an overnight 25 degreesC isopropanol precipitation step. The purified DNA has excellent spectral qualities, is efficiently digested by restriction endonucleases, and is suitable for long-fragment PCR amplification. (C) 2003 Elsevier Science (USA). All rights reserved.