| 欧景莉,覃剑峰,余炳宁,黄小江,陆祖双,朱鹏锦,陈豪军.杨桃SCoT标记PCR反应体系建立与验证[J].中国南方果树,2015,44(3): |
| 杨桃SCoT标记PCR反应体系建立与验证 |
| Establishment and verification of SCoT Amplification System for Averrhoa carambola |
| 投稿时间：2015-01-20 修订日期：2015-03-10 |
| DOI： |
| 中文关键词: 杨桃，SCoT，单因素试验，体系优化 |
| 英文关键词:Averrhoa carambola SCoT Single factor experiment System optimization |
| 基金项目:农业部农垦局物种资源保护财政专项经费；广西壮族自治区直属公益性科研院所基本科研业务费专项 |
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| 中文摘要: |
| 用单因素设计法对影响杨桃SCoT-PCR反应体系的主要因素Mg2+, dNTPs,引物, Taq DNA 聚合酶及DNA 模板浓度进行优化。结果表明，适于杨桃研究的SCoT-PCR最佳反应体系为：总体积为20μL的反应体系中，含2.5mmol/L Mg2+，0.3mmol/L dNTPs，30mg/L模板DNA，1.00μmol/L引物和0.4U Taq DNA聚合酶。用不同引物及杨桃DNA对该体系进行验证，扩增条带清晰，结果稳定可靠，证明该反应体系适用于杨桃SCoT-PCR扩增。 |
| 英文摘要: |
| SCoT-PCR amplification system for Averrhoa carambola was optimized based on single factor experiment design, such as Mg2+, dNTPs, DNA template, Taq DNA polymerase and primer. The results showed that the optimum concentrations for SCoT-PCR amplification system were 2.50 mmol/L Mg2+, 0.3 mmol/L dNTPs, 30 mg/L template DNA, 1.00 μmol/L primer, 0.4 U Taq DNA polymerase, and 2μL 10×buffer in total 20 μL reaction system, respectively. The optimum SCoT-PCR system was tested by different primers and carambola germplasms, obtaining steady, reliable and high resolution bands. It was suggested the reaction system was suitable for the carambola SCoT-PCR amplification.amplification. |
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