| 柳杉CAPS 遗传标记反应体系的优化 |
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| 引用本文： | 杨萍,郑前剑,赵明水,汤定钦. 柳杉CAPS 遗传标记反应体系的优化[J]. 浙江农林大学学报, 2006, 23(1): 52-55 |
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| 作者姓名： | 杨萍 郑前剑 赵明水 汤定钦 |
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| 作者单位： | 1.浙江林学院 浙江省现代森林培育技术重点实验室, 浙江 临安 311300;2.浙江省天目山国家级自然保护区管理局, 浙江 临安 311300 |
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| 摘 要： | 建立柳杉Cryptomeria fortunei CAPS 反应优化体系是进行柳杉遗传多样性研究的前提。通过对影响柳杉CAPS 遗传标记反应体系结果主要因子的研究, 确定了最佳的柳杉CAPS 扩增反应体系:含有2.0 L 10 Buffer , 0.08 mmolL-1dNTPs , 200mgL-1铸型DNA , 1.5 mmolL-1MgCl2 , 0.1 molL-1引物和41.675 nkat Taq DNA 聚合酶的20 L 反应液。扩增程序是94℃预变性5min , 然后94 ℃变性40 s , 58 ℃退火40 s , 72 ℃延伸80 s , 共35 个循环, 然后在72 ℃延伸5 min 。PCR 产物10.0 L, 加入2.0 L 10 Buffer , 83.350 nkat 内切酶, 7.5 L 双蒸水构成20 L 酶切反应体系。图5 表1 参8
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| 关 键 词： | 植物学 柳杉 切割的扩增产物多态性序列(CAPS) DNA 标记 反应体系 优化 |
| 收稿时间： | 2004-11-16 |
Optimization of the reaction system for CAPS marker analysis in Cryptomeria fortunei |
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| YANG Ping,ZHENG Qian-jian,ZHAO Ming-shui,TANG Ding-qin. Optimization of the reaction system for CAPS marker analysis in Cryptomeria fortunei[J]. Journal of Zhejiang A&F University, 2006, 23(1): 52-55 |
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| Authors: | YANG Ping ZHENG Qian-jian ZHAO Ming-shui TANG Ding-qin |
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| Affiliation: | 1.Key Labratory for Modern Silvicultural Technology of Zhejiang Province , Zhejiang Forestry College , Lin’an 311300, Zhejiang , China;2..Management Office , National Nature Reserve of Mount Tianmu , Lin’an 311300 , Zhejiang , China |
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| Abstract: | An optimized system for CAPS reaction in Cryptomeria fortunei is a precondition for analyzing genetic diversity of Cryptomeria fortunei .The authors optimized a reaction system for CAPS in Cryptomeria fortunei by analyzing different factors .The results showed that the best one was 20 L amplification reaction solution composed of 2.0 L 10 Buffer , 0.08 mmolL-1dNTPs , 200 mgL-1 template DNA , 1.5 mmolL-1 Mg2+ , 0.1 mol L-1primer and 41.675 nkat Taq DNA polymerase , which was amplified under the program of predenaturing at 94 ℃ for 5min , followed by 35 cycles of denaturing at 94 ℃for 40 s , annealing 58 ℃for 40 s , extension 72 ℃for 80 s , and a final extension 72 ℃ for 5 min .The restriction digestion system consisted of 10 L PCR products , 2.0 L 10 Buffer , 83.350 nkat restriction enzyme and 7.5 L ddH2O .[ Ch , 5 fig .1 tab .8 ref .] |
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