| 刺槐AFLP分析体系的建立与优化 |
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| 引用本文： | 籍新波,张连成,韩宏伟,张军,王进茂.刺槐AFLP分析体系的建立与优化[J].河北林果研究,2012,27(2):139-142. |
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| 作者姓名： | 籍新波 张连成 韩宏伟 张军 王进茂 |
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| 作者单位： | 1. 河北农业大学林学院,河北保定,071000
2. 天津滨海新区塘沽林业工作站,天津,300450 |
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| 基金项目： | 国家林业公益性行业科研专项 |
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| 摘 要： | 以10份刺槐种质资源为试材,通过对影响AFLP反应体系关键因素的优化,建立了刺槐AFLP分析的反应体系,即15ng/μL模板DNA在37℃下双酶切(MseI、EcoRI各0.15U/μL)2h,0.075U/μL T4DNA连接酶连接12h,连接产物、预扩增产物稀释10倍后扩增效果最佳。并利用该体系筛选出引物E+3/M+3,构建了刺槐种质资源的AFLP指纹图谱。
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| 关 键 词： | 刺槐 AFLP 分析体系 |
Establishment and optimization of AFLP reaction system in Robinia pseudoacacia |
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| JI Xin-bo
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ZHANG Lian-cheng
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HAN Hong-wei
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ZHANG Jun
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WANG Jin-mao.Establishment and optimization of AFLP reaction system in Robinia pseudoacacia[J].Hebei Journal of Forestry and Orchard Research,2012,27(2):139-142. |
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| Authors: | JI Xin-bo ZHANG Lian-cheng HAN Hong-wei ZHANG Jun WANG Jin-mao |
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| Institution: | 1 Department of Forestry, Agricultural University of Hebei ,Baoding 071000, China 2 Forestry Workstation of Tanggu, Tianjin Binhai New Area, Tianjin 300450, China) |
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| Abstract: | AFLP procedure used for Robinia pseudoacacia DNA fingerprinting was estab-lished by analyzing some critical factors. Some improved factors were below： template prod-ucts DNA （15 ng/μL） was digested by MseI and EcoRI （MseI and EcoRI each 0. 15 U/pL） at 37℃ for 2 h, ; digestion was ligated by T4DNA ligase （0. 075 U/μL） for 12 hours; the product of ligation were diluted to 10 times for pre-amplification; the products of pre-amplifi-cation were diluted to 10 times for selective amplification. The fingerprinting map of Robinia pseudoacacia germplasm was constructed with primer E＋3 /M＋3. |
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| Keywords: | Robinia pseudoacacia AFLP reaction system |
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