| 摘要: |
| 通过单因素试验，对杂种松SRAP-PCR反应体系的各主要影响因子(Mg2+浓度、dNTP浓度、引物浓度以及Taq DNA 聚合酶用量)进行了优化筛选，最终得出杂种松SRAP-PCR反应的最佳体系为：25ul体系中含有Mg2+ 2.0 mmol/L、dNTPs 0.25mmol/L、Taq 酶1.0U，引物0.4 umol/L，模板DNA 30-50ng以及1%体积的DMSO。采用该优化体系，对两种杂种松（火加松、湿加松）基因组DNA进行SRAP扩增，表明大部分引物对扩增出来的带清晰可辨、背景较少，完全满足分子标记的要求，为杂种松SRAP标记的相关研究奠定了基础。 |
| 关键词: 杂种松 SRAP 反应体系优化 |
| DOI： |
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| Optimization of SRAP-PCR Reaction System in Hybrid Pines |
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Luo Rui,Yang Huixiao,Huang Shaowei,Zhao Fencheng,Liu Tianyi,Liu Chunxin
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College of Forestry, South China Agricultural University
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| Abstract: |
| Through single-factor test, concentrations of the main factors influencing SRAP-PCR reaction(Mg2+, dNTP, primer and Taq DNA polymerase, respectively ) in two pine hybrids were optimized. The optimized reaction system was that a total 25ul containing 1×PCR Buffer, Mg2+ 2.0 mmol/L, dNTPs 0.2 5mmol/L, Taq DNA polymerase 1.0U, primer 0.4 umol/L, template DNA 30-50ng and 1% DMSO. Using this optimized system, SRAP-PCR reactions were performed on two hybrid pines(F1 of slash pine×Caribbean pine and loblolly pine×Caribbean pine), and scorable bands were amplified. This study laid a solid foundation for further researches on SRAP markers in hybrid pines. |
| Key words: hybrid pine SRAP reaction system optimization |
