| 萱草ISSR-PCR最佳反应体系的建立 |
| |
| 引用本文： | 黎海利,董丽,谭飞理.萱草ISSR-PCR最佳反应体系的建立[J].安徽农业科学,2008,36(12):4884-4885. |
| |
| 作者姓名： | 黎海利 董丽 谭飞理 |
| |
| 作者单位： | 北京林业大学国家花卉工程研究中心,北京,100083;北京林业大学国家花卉工程研究中心,北京,100083;北京林业大学国家花卉工程研究中心,北京,100083 |
| |
| 基金项目： | 国家林业局948项目[2002-08(2)]. |
| |
| 摘 要： | 目的]为利用ISSR标记研究萱草的遗传多样性和和辅助育种奠定基础。方法]从萱草中提取基因组DNA,建立萱草的ISSR-PCR反应体系,并通过正交试验对其进行优化。结果]萱草的最佳ISSR-PCR反应体系为:ddH2O16.8μl、2.5 mmol/LdNTP2.0μl、10×buffer 2.5μl、10μmol/LPrimer0.3μl、50 ng/LDNA3μl、Taq酶1 U,总体积25μl。萱草的最佳ISSR扩增反应程序为:94℃变性5 min,94℃变性45 s、48.3℃退火1 min、72℃延伸2 min,共38个循环,最后72℃延伸7 min。该反应程序下进行的ISSR扩增,扩增产物最多,银染的效果最好。结论]该研究建立了萱草ISSR-PCR的最佳反应体系和反应程序,通过ISSR扩增可获得清晰、稳定的条带。
|
| 关 键 词： | 萱草 ISSR 最佳反应体系 |
| 文章编号： | 0517-6611(2008)12-04884-02 |
| 修稿时间： | 2008年2月27日 |
Establishment of the Optimum Reaction System for ISSR-PCR of Hemerocallis fulva L. |
| |
| LI Hai-li.Establishment of the Optimum Reaction System for ISSR-PCR of Hemerocallis fulva L.[J].Journal of Anhui Agricultural Sciences,2008,36(12):4884-4885. |
| |
| Authors: | LI Hai-li |
| |
| Abstract: | Objective] The research aimed to lay the foundation for studying the genetic diversity and the assistant breeding of Hemerocallis fulva L.by using ISSR markers.Method] The genomic DNA was extracted from H.fulva.ISSR-PCR reaction system of H.fulva was established and optimized in the orthogonal test.Result] The optimum reaction system for ISSR-PCR of H.fulva was as follows: ddH2O 16.8 μl,2.5 mmol/L dNTP 2.0 μl,10×buffer 2.5 μl,10 μmol/L Primer 0.3 μl,50 ng/L DNA 3 μl,Taq enzyme 1U,with the total volume of 25 ul.The optimum reaction procedure for ISSR amplification of H.fulva was as follows: denaturation for 5 min at 94 ℃,30 cycles of denaturation for 45 s at 94 ℃,anneal for 1 min at 48.3 ℃、extension for 2 min at 72 ℃,finally extension for 7 min at 72 ℃.Through ISSR amplification under this reaction procedure,the amplified products were most and the sliver-staining effect was best.Conclusion] The optimum reaction system and reaction procedure for ISSR-PCR of H.fulva was established in this research and clear and stable bands could be obtained though ISSR amplification. |
| |
| Keywords: | Hemerocallis fulva L ISSR Optimum reaction system |
| 本文献已被 CNKI 万方数据 等数据库收录！ |
|
