|
|||||
|
|
| 苦楝RAPD反应体系的优化 | |
| 引用本文： | 陈羡德,陈礼光,陈珺,罗增炎,张志欣,郑郁善.苦楝RAPD反应体系的优化[J].西南林学院学报,2008,28(1):43-47. |
| 作者姓名： | 陈羡德 陈礼光 陈珺 罗增炎 张志欣 郑郁善 |
| 作者单位： | 1. 福建农林大学林学院,福建,福州,350002 2. 漳州师范学院生物系,福建,漳州,363000 3. 连城县林业局,福建,连城,366200 |
| 基金项目： | 福建省科技厅重点资助项目(2005N003). |
| 摘 要： | 以苦楝叶片提取的基因组DNA为材料,通过单因素多水平梯度试验,筛选DNA模板,Mg^2＋,Taq DNA聚合酶,dNTPs和随机引物的浓度及用量,建立苦楝RAPD技术分析体系.结果表明：当基因组DNA浓度为60 ng/μL,镁离子浓度为3.0 mmol/L,dNTP浓度为0.25 mmol/L,引物浓度为0.30μmol/L,Taq DNA聚合酶用量为1 U/20μL,反应体系总体积20μL时,出现可辨认的清晰谱带.其扩增程序为：94℃预变性2 m in;然后38个循环（94℃变性30 s,37℃退火1 min,72℃延伸80 s）;最后72℃延伸8 min,4℃保存. |
| 关 键 词： | 苦楝 RAPD反应 体系优化 |
| 文章编号： | 1003-7179(2008)01-0043-05 |
| 修稿时间： | 2007年11月23 |
Optimization of Melia azedarach RAPD Reaction System |
|
| CHEN Xian-de,CHEN Li-guang,CHEN Jun,LUO Zeng-yan,ZHANG Zhi-xin,ZHENG Yu-shan.Optimization of Melia azedarach RAPD Reaction System[J].Journal of Southwest Forestry College,2008,28(1):43-47. | |
| Authors: | CHEN Xian-de CHEN Li-guang CHEN Jun LUO Zeng-yan ZHANG Zhi-xin ZHENG Yu-shan |
| Institution: | CHEN Xian-de, CHEN Li-guang , CHEN Jun , LUO Zeng-yan , ZHANG Zhi-xin , ZHENG Yu-shan (1. Forestry College, Fujian Agriculture and Forestry University, Fuzhou Fujian 350002, China; 2. Department of Biology, Zhangzhou Normal College, Zhangzhou Fujian 363000, China; 3. Forestry Bureau of Liancheng County, Liancheng Fujian 366200, China) |
| Abstract: | Taking genomic DNA extracted from Melia azedarach foliage as experimental material, a RAPD analysis system was established by means of single factor multi - level gradient experiment to select suitable concentration and dosage for template, Mg^2＋ , Taq DNA polymerase, dNTPs, and the random primers. The results showed that： Clear electrophoresis bands could be obtained with 60 ng/μL concentration of genomic DNA, 3.0 mmol/L of Mg^2＋ , 0.25 mmol/L of dNTP, 0.30 μmol/L of primer, 1 U/（20 μL） of Taq DNA polymerase and when the total reaction volume was 20 μL. The optimum RAPD program was 2 min predenaturation at 94℃, followed by 38 reaction cycles, 8 min extension at 72 ℃ and final storage at 4 ℃. Each reaction cycle included 30 sec of denaturation at 94 ℃, 1 min annealing at 37℃ and 80 sec extension at 72 ℃. |
| Keywords: | Melia azedarach RAPD reaction system optimization |
| 本文献已被 维普 万方数据 等数据库收录！ | |