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| 大花君子兰ISSR-PCR反应体系的建立与优化 | |
| 引用本文： | 庞赫,郭太君,胡志军,潘肃.大花君子兰ISSR-PCR反应体系的建立与优化[J].吉林农业大学学报,2010,32(2):167-171. |
| 作者姓名： | 庞赫 郭太君 胡志军 潘肃 |
| 作者单位： | 1. 吉林农业大学园艺学院,长春,130118 2. 长春市幸福乡红嘴子村君子兰基地,长春,130000 3. 集安市人参特产业办公室,集安,134200 |
| 基金项目： | 吉林省教育厅“十一五”科学技术研究项目[(2007)第70号] |
| 摘 要： | 以大花君子兰为材料研究了PCR反应体系的主要成分及退火温度对君子兰ISSR扩增结果的影响,并对影响PCR反应体系的主要成分及退火温度进行筛选和优化,确立了适合君子兰ISSR-PCR分析的最佳反应体系:在20μL的反应体系中,Mg2+为2.0 mmol/L,模板DNA用量为50 ng/μL,Taq酶0.4 U,dNTPs浓度为0.15 mmol/L,引物浓度为0.4 mmol/L。筛选出了13个适宜君子兰遗传分析的ISSR引物,确定最适宜退火温度为52~56℃。 |
| 关 键 词： | 君子兰 ISSR 反应体系 优化 |
| 收稿时间： | 2009-08-25 |
| 修稿时间： | 2009-10-21 |
Establishment and Optimization of ISSR-PCR Reaction System in Clivia miniata |
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| PＡＮＧ He,GUO Tai-jun,HＵ Zhi-jun|PAN Su.Establishment and Optimization of ISSR-PCR Reaction System in Clivia miniata[J].Journal of Jilin Agricultural University,2010,32(2):167-171. | |
| Authors: | PＡＮＧ He GUO Tai-jun HＵ Zhi-jun|PAN Su |
| Institution: | 1.College of Horticulture| Jilin Agricultural University, Changchun 130118, China;2. Hongzuizi Village Clivia Base of Xingfu Town of Changchun City, Changchun 130000,China； ３Ji′an Ginseng Special Industry Office,Ji′an 134200,China |
| Abstract: | The factors affecting the ISSR (inter-simple sequence repeat) result of Clivia (Clivia miniata) were studied.By adjusting Mg~(2+),dNTPs,primer,Taq DNA polymerase and annealing temperature were optimized.The optimal ISSR-PCR reaction conditions for Clivia miniata were determined as follows,in 20 μL PCR reaction system,2.0 mmol/L Mg~(2+),50 ng/μL template DNA,0.4 U Taq DNA polymerase,0.15 mmol/L dNTPs,0.4 μmol/L primer.Thirteen ISSR primers that are suitable for Clivia were screened,and the optimal annealing temperature for the ISSR reaction was 52-56℃. |
| Keywords: | Clivia miniata ISSR reaction system optimization |
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