| 喙尾琵甲DNA提取方法比较与AFLP反应体系的建立 |
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| 引用本文： | 赵 敏,冯 颖,和 锐,陈晓鸣,冀焕红. 喙尾琵甲DNA提取方法比较与AFLP反应体系的建立[J]. 林业科学研究, 2010, 23(2): 234-240 |
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| 作者姓名： | 赵 敏 冯 颖 和 锐 陈晓鸣 冀焕红 |
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| 作者单位： | 中国林业科学研究院资源昆虫研究所,国家林业局资源昆虫培育与利用重点实验室,云南,昆明,650224 |
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| 基金项目： | 国家林业局林业科学技术重点研究项目(2006-65);国家科技支撑计划课题(2006BAD06B07) |
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| 摘 要： | 采用CTAB法、改进SDS-蛋白酶K法、SDS-饱和氯化钠法和提取试剂盒对喙尾琵甲的肌肉、虫卵和幼虫进行DNA提取,比较了DNA的提取和保存质量,并对AFLP试验的酶切时间、预扩增产物稀释倍数和选择性扩增引物量等关键因子进行试验和优化。结果表明:4种方法对肌肉组织提取的DNA质量都较好;3种传统方法提取的DNA的保存时间长于试剂盒提取的DNA;使用EcoR Ⅰ/Mse Ⅰ内切酶组合,10 U EcoR Ⅰ和2 U Mse Ⅰ分两步各酶切2 h,T4连接酶3 h连接后,以20倍稀释的预扩增产物和5 ng/35 ng的E+3/M+3引物量进行选择性扩增,建立了喙尾琵甲AFLP分子标记体系。
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| 关 键 词： | 喙尾琵甲 基因组DNA DNA提取 AFLP |
Genomic DNA Extraction Method and AFLP Amplification Reaction System of Blaps rhynchopetera (Coleoptera: Tenebrionidae) |
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| ZHAO Min,FENG Ying,HE Rui,CHEN Xiao-ming and JI Huan-hong. Genomic DNA Extraction Method and AFLP Amplification Reaction System of Blaps rhynchopetera (Coleoptera: Tenebrionidae)[J]. Forest Research, 2010, 23(2): 234-240 |
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| Authors: | ZHAO Min FENG Ying HE Rui CHEN Xiao-ming JI Huan-hong |
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| Affiliation: | Research Institute of Resource Insects/a>;Chinese Academy of Forestry/a>;Key Laboratory of Cultivation and Utilization of Resource Insects ofState Forestry Administration/a>;Kunming 650224/a>;Yunnan/a>;China |
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| Abstract: | The study compared the quality of DNA extracting from muscles,eggs and larvae of Blaps rhynchopetera with CTAB method,improved SDS-protease-K method,SDS-NaCl method and DNA extraction Kit.Some key factors affecting the time of digestion,dilution of pre-amplification product and the amount of selective amplification primer were studied and an optimized AFLP reaction system of B.rhynchopetera was established.High quality genomic DNA could be isolated using four methods from this beetle muscles.DNA extracted w... |
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| Keywords: | Blaps rhynchopetera genomic DNA DNA extraction AFLP |
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