| 湿地松、加勒比松SRAP反应体系的优化及引物筛选 |
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| 引用本文： | 李义良.湿地松、加勒比松SRAP反应体系的优化及引物筛选[J].广东林业科技,2011,27(3):8-13. |
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| 作者姓名： | 李义良 |
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| 作者单位： | 广东省林业科学研究院 广东广州510520 |
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| 基金项目： | 国家林业公益性行业科研专项（201104014），广东省林业科技创新专项资金项目（2008KJCX00501，2009KJCX00701），广东省科技厅项目（2007A0202000011），广东省林业局林业种苗与基地管理总站项目（2009），北京林业大学林木育种国家工程实验室开放课题项目(FOP20103)。 |
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| 摘 要： | 采用正交设计L9(34)对影响湿地松、加勒比松SRAP反应体系的DNA浓度、dNTPs浓度、引物用量、Taq DNA聚合酶浓度4个因素进行了优化试验,结果表明:扩增反应体系5优于其他体系,即25μL体系中含有DNA 40 ng、dNTPs 1mmol/L、引物10 μmol/L、DNA聚合酶1.0U.在此基础上,通过单...
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| 关 键 词： | 湿地松 加勒比松 SRAP-PCR 体系优化 引物筛选 |
| 收稿时间： | 3/1/2011 12:00:00 AM |
| 修稿时间： | 2011/6/10 0:00:00 |
Optimization of SRAP Reaction System and Primers Screening of Pinus elliottii and P.caribaea |
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| zhaofencheng.Optimization of SRAP Reaction System and Primers Screening of Pinus elliottii and P.caribaea[J].Forestry Science and Technology of Guangdong Province,2011,27(3):8-13. |
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| Authors: | zhaofencheng |
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| Institution: | Li Yiliang Zhao Fencheng Wu Huishan(Guangdong Academy of Forestry,Guangzhou,510520) |
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| Abstract: | The orthogonal design,L9(34) included 4 factors(DNA template,dNTPs,primers and Taq polymerase) and 3 levels was used to optimize SRAP reaction system for Pinus elliottii and P.caribaea.The results showed that: PCR reaction system 5 is superior to other systems,that is,the system contains 25 μL,DNA 40 ng,dNTPs 1 mmol/L,primer 10 μmol/L,DNA polymerase 1.0 U.Based on the results of experiment from orthogonal design,the proper conditions of the key factors affecting the stability of SRAP system were determine by single-factor completely randomized design.Each 25 μL PCR reaction mixture consisted of 40 ng of genomic DNA,1 mmol/L of dNTPs,5 μM of primer and 1.0 unit of Taq polymerase.Total of 216 primer pairs were used in SRAP amplification and 42 combinations were selected because of their clear banding patterns,consistent amplifications and good polymorphism.The optimal SRAP reaction system was established,which would provide the basis for evaluation of genetic diversity,moleular marker assisted breeding and linkage mapping for the P.elliottii and P.caribaea.; |
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| Keywords: | Pinus elliottii P caribaea SRAP-PCR optimization of system primers screening |
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