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木麻黄AFLP分析体系的建立及引物筛选
引用本文:黄桂华,高丽丽,仲崇禄,苏晓华,张勇,陈羽,陈珍,谢冬勇.木麻黄AFLP分析体系的建立及引物筛选[J].中南林业科技大学学报,2007,27(1):20-24,34.
作者姓名:黄桂华  高丽丽  仲崇禄  苏晓华  张勇  陈羽  陈珍  谢冬勇
作者单位:中国林业科学研究院热带林业研究所 广东广州510520(黄桂华,仲崇禄,张勇,陈羽,陈珍),华南农业大学园艺学院 广东广州510642(高丽丽),中国林业科学研究院林业研究所 北京100091(苏晓华),广州市绿化公司 广东广州510170(谢冬勇)
基金项目:国家高技术研究发展计划(863计划);广东省自然科学基金;中国-澳大利亚合作项目;广东省科技计划
摘    要:采用CTAB法改良技术提取木麻黄基因组DNA,在提取液中加入2%β-巯基乙醇防氧化和2%的PVP去除酚类等物质,获得高质量的基因组DNA,以满足AFLP对模板DNA高质量的要求;采用分步酶切和连接后,进行预扩和选扩,摸索建立和优化了木麻黄AFLP分析体系,探讨了木麻黄AFLP分析的影响因素;从64对AFLP核心引物组中筛选出了8对适合木麻黄分析的引物组合.同时探讨了AFLP技术在木麻黄研究领域的应用前景.

关 键 词:生物技术  分析体系  引物  木麻黄
文章编号:1000-2502(2007)01-0020-05
收稿时间:2006-09-10
修稿时间:2006-09-10

Establishment of the Analytical System for Casuarinaceae AFLP and Its Primer Selection
HUANG Gui-hua,GAO Li-li,ZHONG Chong-lu,SU Xiao-hua,ZHANG Yong,CHEN Yu,CHEN Zhen,XIE Dong-yong.Establishment of the Analytical System for Casuarinaceae AFLP and Its Primer Selection[J].Journal of Central South Forestry University,2007,27(1):20-24,34.
Authors:HUANG Gui-hua  GAO Li-li  ZHONG Chong-lu  SU Xiao-hua  ZHANG Yong  CHEN Yu  CHEN Zhen  XIE Dong-yong
Institution:1. Research Institute of Tropical Forestry, CAF, Guangzhou 510520, Guangdong, China; 2. South China Agricultural University, Guangzhou 510641, Guangdong, China; 3. Institute of Forestry Research, CAF, Beijing 100091, China; 4. Guangzhou Green Company, Guangzhou 510170, Guangdong, China
Abstract:An improved method of CTAB was used in this study to extract the total DNA of Casuarinaceae.Then,high quality genomic DNA was isolated for AFLP analysis with 2% of mercapto-ethanol in the extracting solution to prevent formation of oxides and 2% of PVP to eliminate phenols from genomic DNA.By means of double enzyme restriction,adapter-ligation,pre-amplification and selective amplification,AFLP analytical system of Casuarinaceae was established and optimized.8 primer combinations were selected from 64 pairs of core combinations of AFLP after a series of tests.The application prospect of AFLP in Casuarinaceae research field was also discussed.
Keywords:AFLP
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