| 姚婧，唐燕琼，黄明忠，丁亚操，杨光穗.火焰兰属SRAP-PCR 体系的建立优化及验证[J].广东农业科学,2014,41(20):141-144 |
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火焰兰属SRAP-PCR 体系的建立优化及验证 |
| Optimization and verification for SRAP - PCR system in Renanthera Loureiro |
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| DOI： |
| 中文关键词: 火焰兰 SRAP-PCR 体系优化 |
| 英文关键词: Renanthera Loureiro SRAP-PCR system optimization |
| 基金项目:教育部热带作物新品种选育工程中心与作物学重点学科联合资助项目（lhxm-2013-7）；中国热科院热带作物品种资源研究所基本科研业务费专项重点项目（1630032013035） |
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| 中文摘要: |
| 为建立火焰兰SRAP-PCR 体系，采用U20(55)均匀试验设计，，以10 份火焰兰种质基因组DNA 为模板，用3对SRAP引物对DNA模板、Mg2+、引物、TaqDNA聚合酶和dNTPs等5个因素的浓度进行优化组合和验证。结果表明，提取的火焰兰基因组DNA纯度高、完整性好；利用引物对Me6/Em6 进行PCR 扩增，对20个处理初步筛选出扩增条带较清晰，多态性丰富的处理，再用2个DNA模板进行复筛，获得最佳反应体系（20μL）：模板DNA60ng、Mg2+2.0 mmol/L、引物1.0μmol/L、Taq DNA聚合酶1.2 U、dNTPs 0.20 mmol/L。最后用2对引物、8份火焰兰种质来验证优化SRAP-PCR 体系，其所获条带清晰，多态性丰富。 |
| 英文摘要: |
| In order to optimize SRAP-PCR reaction system for Renanthera Loureiro, the U20(55) uniform experimental design was applied to optimize concentration of DNA template, Mg2+, primers, Taq DNA polymerase and dNTPs. Then the system was verified with 10 R. germplasm genome DNA as template and three pairs of SRAP primers. The results showed that the extraction of genomic DNA was of high purity and had good integrity; The bands of 20 treatments amplifing with Me6/Em6 primers, were clear, polymorphic processing of rich; Further screening using 2 DNA template for getting, the best system was the total volume of 20 μL reaction solution containing 60 ng genomic templates DNA, 2.0 mmol/L Mg2+, 1.0 μmol/L Primers, 1.2 U Taq DNA polymerase, 0.20 mmol/L dNTP. The bands obtained in the verification experiment were clear and highly polymorphic. |
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