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| 山茶ISSR扩增条件的优化 | |
| 引用本文： | 谈探,金则新,李钧敏,潘仙鹏.山茶ISSR扩增条件的优化[J].福建林业科技,2007,34(2):24-27. |
| 作者姓名： | 谈探 金则新 李钧敏 潘仙鹏 |
| 作者单位： | 1. 北京林业大学自然保护区学院,北京,100083;台州学院生态研究所,浙江,临海,317000 2. 台州学院生态研究所,浙江,临海,317000 3. 台州市农业科学研究院,浙江,临海,317000 |
| 摘 要： | 以山茶基因组DNA为材料,测试山茶ISSR扩增的最适退火温度,并采用单因素试验,测试了模板DNA量,Mg2+浓度,dNTP浓度,BSA浓度,引物用量,Taq酶用量等6个因素对山茶ISSR扩增的影响。结果显示:山茶ISSR扩增的最适退火温度为56.3℃;适宜的扩增体系为:10μL PCR反应体积中,1×Taq酶配套缓冲液(10 mmol.L-1Tris.HCl pH 9.0,50mmol.L-1KCl,0.1%Triton X-100),2 mmol.L-1MgCl2,0.6 mmol.L-14×dNTP,2 mg.ml-1BSA,16 ng模板DNA,10 pmol引物,0.5 U Taq酶。 |
| 关 键 词： | 山茶 ISSR 扩增条件 优化 |
| 文章编号： | 1002-7351（2007）02-0024-04 |
| 修稿时间： | 2006-09-27 |
Optimization of the ISSR reaction system of Camellia japonica |
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| TAN Tan,JIN Ze-xin,LI Jun-min,PAN Xian-peng.Optimization of the ISSR reaction system of Camellia japonica[J].Journal of Fujian Forestry Science and Technology,2007,34(2):24-27. | |
| Authors: | TAN Tan JIN Ze-xin LI Jun-min PAN Xian-peng |
| Institution: | 1. College of Natural Reserves, Beijing Forestry University, Beijing 100083, China; 2. Institute of Ecology, Taizhou University, Linhai, Zhejiang 317000, China; 3. Taizhou Academy of Agricultural Sciences, Linhai, Zhejiang 317000, China |
| Abstract: | The suitable ISSR-PCR amplification condition of the genomic DNA of Camellia japonica was optimized and the optimal annealing temperature was determined.Using single factor test method,the effects of 6 factors, such as Mg2+ concentration,dNTP concentration,BSA dosage,DNA templates dosage,primer dosage and Taq DNA polymerase dosage for ISSR amplification were tested.The optimal annealing temperature was 56.3℃ and the optimal ISSR reaction system for Camellia japonica was as follows: 1×Taq polymerase buffer 10 mmol·L-1 Tris·HCl(pH 9.0), 50 mmol·L-1 KCl,0.1% Triton X-100],2 mmol·L-1 MgCl2,0.6 mmol·L-1 4×dNTP,2 mg·ml-1 BSA,16 ng DNA template,10 pmol primer,0.5 U Taq DNA polymerase in total 10 μL reaction volume. |
| Keywords: | Camellia japonica ISSR reaction system optimization |
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