| 摘要: |
| 以刨花润楠（Machilus pauhoi）1.5 a 生小苗幼嫩叶片为试材，对影响刨花润楠 SRAP-PCR 扩
增的模板 DNA 量、引物、dNTP 和 Mg 2 体积摩尔浓度、Taq DNA 聚合酶、退火温度 6 个主要因素进
行优化。结果表明，SRAP-PCR 的最佳反应体系为：25 μL 的 SRAP-PCR 反应体系中，2.5 μL 10×PCR
buffer、模板 DNA 量 60 ng、Mg 2 2.0 mmol/L、dNTP 0.225 mmol/L、引物 0.3 μmol/L 和 Taq DNA 聚合酶
1.25 U。对优化的反应体系和扩增程序的验证结果表明，优化的刨花润楠 SRAP-PCR 反应体系和扩增程
序是稳定可行的。 |
| 关键词: 刨花润楠 SRAP-PCR 体系优化 |
| DOI： |
| 分类号:S718.43; S792.23 |
| 基金项目:“十二五”国家科技支撑项目；广东省林业科技创新项目 |
|
| Establishment and Optimization of SRAP-PCR System for Machilus pauhoi |
|
zhoupeng,linwei,zhouxiangbin and chenxiaoyang
|
|
Guangdong Eco-Engineering Polytechnic,College of Forestry and Landscape Architecture, Sourth China Agricultural University,College of Forestry and Landscape Architecture, Sourth China Agricultural University,College of Forestry and Landscape Architecture, Sourth China Agricultural University
|
| Abstract: |
| Machilus pauhoi is a tree specie with variety of economic value and development prospects. This
study aimed to establish an optimized SRAP-PCR system for M. pauhoi, and the young leaves of the 1.5 years
old seedlings were used as test materials. Six quality factors including the template DNA, primer concentration,
dNTP concentration, Mg 2 concentration, Taq DNA polymerase, and annealing temperature were optimized for M.
pauhoi SRAP-PCR assay. The obtained results suggested a optimized reaction system of SRAP-PCR (total 25 μL)
involving 2.5 μL 10×PCR buffer, 60 ng DNA, 2.0 mmol/ L Mg 2 , 0.225 mmol/L dNTP, 0.3 μmol/L primer, 1.25
U Taq DNA polymerase. The verification results showed that the optimized SRAP-PCR reaction system and
amplification program were stale and feasible. |
| Key words: Machilus pauhoi SRAP-PCR system optimization |
