菠萝SRAP反应体系的建立及优化  

Optimization of SRAP System in Pineapple

作　　者：邱文武[1] 孙伟生[2] 窦美安[2] 

机构地区：[1]华南热带农业大学园艺学院,海南儋州571737 [2]中国热带农业科学院南亚热带作物研究所,广东湛江524091

出　　处：《生物技术》2008年第1期39-42,共4页Biotechnology

基　　金：农业部"948"项目(2006G34A);广东省农业攻关计划项目(2005B20901011);中国热带农业科学院基金项目(YKY517)资助

摘　　要：目的:建立一种适合菠萝基因扩增的SRAP反应体系。方法:用改良CTAB法提取菠萝DNA,对扩增结果影响重要的反应组分Taq酶、Mg2+、随机引物及dNTPs进行单因素体系优化,以确定最佳菠萝SRAP反应体系。结果:用这种方法建立的菠萝SRAP反应体系为:20μL反应体系中含1×PCR buffer,2.5mmol/L Mg2+、1.2UTaqDNA聚合酶、0.2mmol/L dNTPs、0.3μmol/L随机引物、20ng DNA模板。结论:用引物Me4-Em4组合对供试菠萝19个品种进行扩增,结果扩增条带清晰、丰富、重复性好,此SRAP反应体系适合菠萝基因型扩增。Objective： A suitable SRAP reaction system for pineapple was established. Methods： In this paper, to extract the pineapple DNA by the revised CTAB method, and to optimize SRAP- PCR amplification system on pineapple in Taq DNA polymerase, Mg^2 ＋ , primer and dNTPs, respectively. To get a suitable PCR system and the result showed that the suitable SRAP reaction system for pineapple was developed： 1X PCR buffer, 2.5mmol/L Mg^2＋ , 1.2U Taq DNA polymerase,0.2mmol/L dNTPs, μmol/L primer and 20ng genomic DNA templates with total 20μL reaction solution. Conclusion： 19 pineapple varieties were amplified by primers Me4 - Em4 combinations, it showed that the bands was clear, rich, good repeatability and the SRAP reaction system was suitable for pineapple.

关 键 词：菠萝 SRAP 体系优化 

分 类 号：Q781[生物学—分子生物学] S688.3[农业科学—观赏园艺]