| 长白山林下参基因组DNA提取及RAPD体系的优化 |
| |
| 引用本文： | 冯薪硕,朴敬淑,吕龙石.长白山林下参基因组DNA提取及RAPD体系的优化[J].延边大学农学学报,2009,31(1):16-20. |
| |
| 作者姓名： | 冯薪硕 朴敬淑 吕龙石 |
| |
| 作者单位： | 1. 延边大学农学院,吉林,龙井,133400
2. 延边州农业技术推广总站,吉林,延吉,133001 |
| |
| 摘 要： | 采用改良的CTAB法提取林下参的基因组DNA，所得的DNA纯度高、质量好，可用于RAPD分析．筛选出的林下参RAPD反应的最佳体系为20“L，反应体系中包括模板DNA20ng，引物20pmol，dNTPs0．1875mmol／L，TaqDNA聚合酶1．5U，Mg^2＋2．0mmol／L，10×Reaction Buffer2．0mmol／L，其余部分用无菌超纯水补充．PCR扩增程序为94℃预变性5min，94℃变性1min，37℃退火1min，72℃延伸2min，40次循环，72℃最终延伸7min．应用优化后的反应体系PCR扩增获得的RAPD指纹图谱带型清晰，重复性好，为通过分子标记获得丰富的林下参遗传信息奠定了基础．
|
| 关 键 词： | 林下参 基因组DNA RAPD体系 |
Optimization of RAPD System and DNA Extraction from Transplanted Wild Ginseng in ChangBai Moutain |
| |
| FENG Xin-shuo,Piao Jing-shu,LU Long-shi.Optimization of RAPD System and DNA Extraction from Transplanted Wild Ginseng in ChangBai Moutain[J].Journal of Agricultural Science Yanbian University,2009,31(1):16-20. |
| |
| Authors: | FENG Xin-shuo Piao Jing-shu LU Long-shi |
| |
| Institution: | FENG Xin-shuo, PIAO Jing-shu, LU Long-shi(1. Agricultural College of Ya nbian University, Longj ing J ilin 133400, China Agricultural Technology Extension Station of Yanbian ,Yanji Jilin 133001,China) |
| |
| Abstract: | High purity and quality DNA extracted of Transplanted wild ginseng was used for RAPD analysis. The best RAPD reaction system optimized was： total volume 20 μL, containing template DNA 20 ng, primer 20 pmol, dNTPs 0. 187 5 mmol/L, MgCI2 2.0 mmol/L, DNA polymerase 1. 5 unit, 10× Reaction Buffer 2.0 mmol/L, completed with ddH20. PCR amplification condition was： pre-denaturation at 94 ℃for 5 min, denaturation at 94 ℃ for 1 min, annealing at 37℃ for 1 rain, extension at 72 ℃ for 2 min, cycle for 40 times. Final extension at 72℃ for 7 rain. RAPD finger printing obtained by the optimized system was clear and repetitive, which provided a certain foundation for getting more extensive genetic information of Transplanted wild ginseng of Changbai Mountain. |
| |
| Keywords: | Transplanted wild ginseng genome DNA RAPD system |
| 本文献已被 维普 万方数据 等数据库收录！ |
|
